Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
DETAILED ACTION
This application is a continuation of 17/336,063, issued as US Patent No. 11,802,279, which is a continuation of 16/226,499, abandoned, which is a continuation of 15/671,086, issued as US Patent No. 10,174,302.
The amendment filed on January 30, 2026 has been entered.
Election/Restrictions
Applicant elected without traverse of Group I with a species election of (1) SEQ ID NO:1 as the L-asparaginase and (2) SEQ ID NO:7 (201 amino acids) as the polypeptide comprising prolines and alanine in the reply filed on August 20, 2024 is acknowledged.
Claims 6, 20 and 25-30 are withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected species/invention, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on September August 20, 2024.
Newly amended claim 6 is withdrawn because claim is directed to non-elected species. The elected species is a fusion protein comprising (1) SEQ ID NO:1 as the L-asparaginase and (2) SEQ ID NO:7 (201 amino acids) or about 200 proline and alanine residues. However, claim 6 is directed to a fusion protein comprising (1) SEQ ID NO:1 as the L-asparaginase and (2) consisting essentially of 400 proline and alanine residues.
Status of Claims
Claims 1, 3-6, 8, and 19-30 are pending.
Claims 6, 20 and 25-30 are withdrawn.
Claims 1, 3-5, 8, 19, and 21-24 are under examination.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on March 11, 2026 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Response to Amendments/Arguments
Claim Rejections - 35 USC § 112(b)
Applicant’s arguments, see page 5 of the Remarks, filed January 30, 2026, with respect to claim 21 have been fully considered and are persuasive. Clam 21 has been amended to replace the indefinite limitation “consist of about” with “consists essentially”. Therefore, the rejection of claim 21 under 35 U.S.C. 112(b) has been withdrawn.
Claim Rejections - 35 USC § 112(a) - Maintained & Amended
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1, 3-5, 8 and 19 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The claims have been broadly interpreted to compass a fusion protein comprising an L-asparaginase comprising or consisting of the amino acid sequence of SEQ ID NO:1 fused to a polypeptide comprising about 200 proline and alanine residues or consisting essentially of 200 proline and alanine residues, wherein the fusion protein has greater asparaginase or glutaminase activity than the unmodified L-asparaginase. Therefore, the claims are directed to a genus of fusion proteins comprising (1) an L-asparaginase having the amino acid sequence of SEQ ID NO:1 and (2) any polypeptide comprising about or consisting essentially of 200 proline and alanine residues, wherein the wherein the fusion protein has greater asparaginase or glutaminase activity than the unmodified L-asparaginase.
MPEP 2163 I. states that to “satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention.
MPEP 2163. II.A.3.(a) sates that “Possession may be shown in many ways. For example, possession may be shown by describing an actual reduction to practice of the claimed invention. Possession may also be shown by a clear depiction of the invention in detailed drawings or in structural chemical formulas which permit a person skilled in the art to clearly recognize that inventor had possession of the claimed invention. An adequate written description of the invention may be shown by any description of sufficient, relevant, identifying characteristics so long as a person skilled in the art would recognize that the inventor had possession of the claimed invention.
According to MPEP 2163.II.A.3.(a).ii), “Satisfactory disclosure of a ‘representative number’ depends on whether one of skill in the art would recognize that the applicant was in possession of the necessary common attributes or features possessed by the members of the genus in view of the species disclosed. For inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus…Instead, the disclosure must adequately reflect the structural diversity of the claimed genus, either through the disclosure of sufficient species that are ‘representative of the full variety or scope of the genus,’ or by the establishment of ‘a reasonable structure-function correlation.’"
The recitation of “fusion protein comprises greater asparaginase or glutaminase activity than the unmodified L-asparaginase” fails to provide a sufficient description of the genus of the fusion protein as it merely describes the functional features of the genus without providing any definition of the structural features of the species within the genus. The specification does not specifically define any of the species that fall within the genus. The specification does not define any structural features commonly possessed by members of the genus that distinguish them from others. One skilled in the art therefore cannot, as one can do with a fully described genus, visualize or recognize the identity of the members of the genus.
Erwinia chrysanthemi L-asparaginase (CAA31239) having 100% sequence identity to the Erwinia chrysanthemi (also known as Chrysnthemi dickeya) L-asparaginase of SEQ ID NO:1 of the instant application was known in the prior art, see Kotzia (J Biotechnol. 2007 Jan 20;127(4):657-69. Epub 2006 Sep 18. – form PTO-1449, copy provided in parent 17/336,063. page 658 and figure 1 on page 661) and P06608 (UniProt Database – form PTO-1449, copy provided in parent application 17/336,063). A polypeptide comprising of proline and alanine residues were known in the prior art, see Skerra (US Patent 9,221,882 – form PTO-1449. Column 50, lines 43-54 and Column 177-178). However, neither the prior art nor the instant specification discloses a fusion protein comprising (1) an L-asparaginase having the amino acid sequence of SEQ ID NO:1 and (2) any polypeptide comprising about or consisting essentially of 200 proline and alanine residues, wherein the wherein the fusion protein has greater asparaginase or glutaminase activity than the unmodified L-asparaginase.
Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – cited previously on form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – cited previously on form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”.
The specification is limited to a fusion protein comprising (1) an L- asparaginase having the amino acid sequence of SEQ ID NO:1 and (2) a polypeptide having the amino acid sequence of SEQ ID NO:7, which is comprised of 201 proline and alanine residues, wherein the wherein the fusion protein has greater asparaginase activity than the unmodified L-asparaginase. While MPEP 2163 acknowledges that in certain situations “one species adequately supports a genus,” it also acknowledges that “[f]or inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus.” In view of the widely variant species encompassed by the genus, the species described above is not enough and does not constitute a representative number of species to describe the whole genus. Therefore, the specification fails to describe a representative species of the claimed genus.
Further one of skill in the art could identify variants of SEQ ID NO:7 or identify polypeptides having about 200 proline and alanine residues. However, there is no teaching regarding which amino acids of SEQ ID NO:7 can be altered nor structural requirements of polypeptides having about 200 proline and alanine residues that when fused to the L-asparaginase of SEQ ID NO:1, the resulting fusion protein has greater asparaginase or glutaminase activity than the unmodified L-asparaginase.
An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function.
Given this lack of description of the representative species encompassed by the genus of the claims, the specification fails to sufficiently describe the claimed invention in such full, clear, concise, and exact terms that a skilled artisan would recognize that applicants were in possession of the inventions of claims 1, 3-5, 8, and 19.
Applicant's arguments filed January 30, 2026 have been fully considered but they are not persuasive. Due to the amendment of claim 21, claims 21-24 are no longer rejected.
Applicant argues that the amended claims meet the written description requirement because a random coil polypeptide substantially lacks a defined secondary and tertiary structure and the specification has provided detailed disclosure on random coil polypeptides consisting solely of proline and alanine residue together with a representative number or species, such as those of SEQ ID NO:7 and SEQ ID NO:9, with no more than 6 identical consecutive amino acid residues. Therefore, applicant argues that it is readily apparent to those skilled in the art that the applicant was in possession of the claimed invention.
This is not found persuasive.
The claims are not directed to a random coil, but a fusion protein comprising the random coil and the L-asparaginase of SEQ ID NO:1, wherein the resulting fusion protein has greater asparaginase activity than the unmodified L-asparaginase. The recitation that the random coil polypeptide has no more than 6 consecutive amino acid residues (which amounts to 3% of the 200 mer) does not provide sufficient description of the random coil polypeptide, such that when fused to the L-asparaginase of SEQ ID NO:1, the resulting fusion protein has greater asparaginase activity than the unmodified L-asparaginase. The specification is limited to a fusion protein comprising (1) an L- asparaginase having the amino acid sequence of SEQ ID NO:1 and (2) a polypeptide having the amino acid sequence of SEQ ID NO:7, which is comprised of 201 proline and alanine residues, wherein the wherein the fusion protein has greater asparaginase activity than the unmodified L-asparaginase. There is no teaching regarding which amino acids of any random coil having about 200 proline and alanine residues having no more than 6 consecutive amino acid residues or (b) the random coil polypeptide having the amino acid sequence of SEQ ID NO:7 can be altered nor structural requirements of polypeptides having about 200 proline and alanine residues that when fused to the L-asparaginase of SEQ ID NO:1, the resulting fusion protein has greater asparaginase or glutaminase activity than the unmodified L-asparaginase.
Hence the rejection is maintained.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 1, 3-5, 8, 19, and 21-24 is/are rejected under 35 U.S.C. 103 as being unpatentable over Skerra (US Patent 9,221,882 – form PTO-1449), Kotzia (J Biotechnol. 2007 Jan 20;127(4):657-69. Epub 2006 Sep 18. – form PTO-1449, copy provided in parent 17/336,063), and P06608 (UniProt Database. 1988. – form PTO-1449, copy provided in parent application 17/336,063).
Regarding claims 1 and 21, Skerra discloses a fusion protein comprising of (1) a biologically active polypeptide, such as an asparaginase, and (2) a random coil polypeptide comprising of proline and alanine residues (abstract, Columns 14-15, Columns 22-23, Columns 31-35 and Table 2 of Column 45, and Column 46). Skerra discloses a random coil polypeptide (PA#1(200)) having the amino acid sequence of SEQ ID NO:61 (comprised in SEQ ID NO:27) consisting of 201 proline and alanine residues (Column 50, lines 43-54 and see the sequence alignment below). Said polypeptide of SEQ ID NO:61 of Skerra has 100% sequence identity to the polypeptide (comprising of prolines and alanines) of SEQ ID NO:7 of the instant application. Said polypeptide of SEQ ID NO:61 of Skerra comprises the amino acid sequence of SEQ ID NO:5 of the instant application (see the sequence alignment below) and has no more than 6 consecutive amino acids that are identical (Column 50, lines 43-54 and see the Column 177-178). Skerra teaches that random coil P/A polypeptides of about 200 alanine and prolines are capable of conferring increased in vivo/in vitro stability, which is useful for biologically active proteins comprising said random coil in medical applications (Column 8, line 59 through Column 9, line 28). Skerra discloses that the random coil P/A polypeptides do not interfere with the biological activity of the biologically active protein and is inert with respect to proteolysis in blood plasma and immunogenicity (Column 26, lines 25-50).
Regarding claims 4 and 22, the random coil polypeptide of Skerra has proline residues that constitute more than 10% and less than 70% of the polypeptide (Column 50, lines 43-54 and Column 177-178).
Regarding claim 5, the random coil polypeptide of Skerra consists essentially of 200 proline and alanine residues (Column 50, lines 43-54 and Column 177-178).
Regarding claims 8 an 23-24, Skerra discloses fusing the random coil polypeptide comprising of alanine and proline residues at the N-terminus or the C-terminus of the biologically active polypeptide/asparaginase (Columns 31-35).
Regarding claim 19, Skerra discloses a pharmaceutical composition comprising said fusion protein (Columns 11-12).
The difference between the method of Skerra and the instant claims is that the method of Skerra does not explicitly discloses a fusion protein comprising the L-asparaginase having 100% sequence identity to SEQ ID NO:1.
However, regarding claims 1, 3, and 21, Kotzia discloses Erwinia chrysanthemi L-asparaginase (CAA31239) having 100% sequence identity to the Erwinia chrysanthemi (also known as Chrysnthemi dickeya) L-asparaginase of SEQ ID NO:1 of the instant application (page 658 and figure 1 on page 661 and see P06608). Kotzia discloses that Erwinia chrysanthemi L-asparaginase is used in treating of acute myeloid leukemia (page 658).
Regarding the property “wherein the fusion protein comprises greater asparaginase .. than the unmodified L-asparaginase” recited in claim 1, such a property is an inherent property of the fusion protein comprising the Erwinia chrysanthemi L-asparaginase fused to the 201 alanine and proline polypeptide (PA#1(200) of SEQ ID NO:61) of Skerra because said fusion protein has identical chemical structure as the fusion protein comprising L-asparaginase of SEQ ID NO:1 of the instant application fused to the 201 alanine and proline polypeptide PA#1(200) of SEQ ID NO:7 of the instant application as described in the specification of the instant application (Examples 5 and 8-9). The fusion protein described in Examples 5 and 8-9 has greater asparaginase activity than the unmodified L-asparaginase. MPEP 2112.01 states that “Products of identical chemical composition can not have mutually exclusive properties.” In re Spada, 911 F.2d 705, 709, 15 USPQ2d 1655, 1658 (Fed. Cir. 1990). A chemical composition and its properties are inseparable.” Since the fusion protein comprising the Erwinia chrysanthemi L-asparaginase fused to the 201 alanine and proline polypeptide (PA#1(200) of SEQ ID NO:61) of Skerra has identical chemical structure as the fusion protein comprising L-asparaginase of SEQ ID NO:1 of the instant application fused to the 201 alanine and proline polypeptide PA#1(200) of SEQ ID NO:7 of the instant application, the recited property “wherein the fusion protein comprises greater asparaginase .. than the unmodified L-asparaginase” is necessarily present and would flow naturally from following the suggestion of the prior art. MPEP 2112 II states that “[t]here is no requirement that a person of ordinary skill in the art would have recognized the inherent disclosure at the relevant time, but only that the subject matter is in fact inherent in the prior art reference.”
Therefore, combining the teachings of Skerra, Kotzia, and P06608, it would have been obvious to one having ordinary skill in the art before the claimed invention was effectively filed to modify the fusion protein of Skerra by substituting the biologically active polypeptide/asparaginase component with Erwinia chrysanthemi L-asparaginase because one of ordinary skill in the art would have been able to carry out such a substitution, and the results were reasonably predictable. The rationale to support a conclusion that the claims would have been obvious is that the substitution of one known element (asparaginase) for another yields predictable results to one of ordinary skill in the art. Therefore, it would have been obvious to one of ordinary skill in the art to replace the prior art biologically active polypeptide/asparaginase with another known and available L-asparaginases, such as the Erwinia chrysanthemi L-asparaginase of Kotzia. One of ordinary skill in the art would have been motivated to modify the fusion protein of Skerra and make a pharmaceutical composition comprising said fusion protein in order to treat acute myeloid leukemia because Kotzia discloses that Erwinia chrysanthemi L-asparaginase is used in treating of acute myeloid leukemia and Skerra discloses that the random coil polypeptide increases the plasma half-life and decreases immunogenicity of the biologically active polypeptide/asparaginase and does not interfere with the biological activity of the biologically active polypeptides. One of ordinary skill in the art would have had a reasonable expectation of success since Skerra discloses a fusion protein comprising of a random coil polypeptide consisting solely of prolines and alanines and biologically active polypeptides, such as an asparaginase, and Kotzia discloses an L-asparaginase.
Therefore, the above references render claims 1, 3-5, 8, 19, and 21-24 prima facie obvious.
Applicant's arguments filed January 30, 2026 have been fully considered but they are not persuasive.
Applicant argues that the claims are not obvious over the cited references because none of the cited reference disclose the property of having greater asparaginase or glutaminase activity. Applicant argues that at the time of the invention, a fusion protein that fuses the Erwinia chrysanthemi L-asparaginase of SEQ ID NO: 1 of Kotzia to the 201 alanine and proline polypeptide of Skerra did NOT exist. Applicant argues that it is well accepted that obviousness cannot be predicated on what is not known at the time an invention is made, even if the inherency of a certain feature is later established. In re Rijckaert, 9 F.2d 1531 (Fed. Cir. 1993). MPEP § 2141.02(V). Applicant argues that the reasonable expectation of success cannot be shown by inherency.
This is not found persuasive.
Due to the amendment of claim 21, claims 21-24 are not directed to a fusion protein wherein the fusion protein has greater asparaginase or glutaminase activity that the L-asparaginase in an unmodified from. Therefore, Applicant’s arguments are moot for claims 21-24.
MPEP 2112. IV. “In relying upon the theory of inherency, the examiner must provide a basis in fact and/or technical reasoning to reasonably support the determination that the allegedly inherent characteristic necessarily flows from the teachings of the applied prior art.”. In the instant case, each component of the fusion protein was known and Skerra provides teachings on assembling the fusion protein. Regarding each component, Skerra discloses a fusion protein comprising of (1) a biologically active polypeptide, such as an asparaginase, and (2) a random coil polypeptide (PA#1(200) comprising of proline and alanine residues, such as the random coil of SEQ ID NO:61, which is identical to the random coil of SEQ ID NO:7 of the instant application (abstract, Columns 14-15, Columns 22-23, Columns 31-35, Table 2 of Column 45, and Column 46, Column 50, lines 43-54 and see the sequence alignment below). Kotzia discloses Erwinia chrysanthemi L-asparaginase (CAA31239) having 100% sequence identity to the Erwinia chrysanthemi (also known as Chrysnthemi dickeya) L-asparaginase of SEQ ID NO:1 of the instant application (page 658 and figure 1 on page 661 and see P06608). The property “wherein the fusion protein comprises greater asparaginase .. than the unmodified L-asparaginase” recited in claim 1 is an inherent property that necessarily flows upon fusing the Erwinia chrysanthemi L-asparaginase to the 201 alanine and proline polypeptide (PA#1(200) of SEQ ID NO:61) of Skerra because said fusion protein has identical chemical structure as the fusion protein comprising L-asparaginase of SEQ ID NO:1 of the instant application fused to the 201 alanine and proline polypeptide PA#1(200) of SEQ ID NO:7 of the instant application as described in the specification of the instant application (Examples 5 and 8-9). The fusion protein described in Examples 5 and 8-9 has greater asparaginase activity than the unmodified L-asparaginase. MPEP 2112.01 states that “Products of identical chemical composition can not have mutually exclusive properties.” In re Spada, 911 F.2d 705, 709, 15 USPQ2d 1655, 1658 (Fed. Cir. 1990). A chemical composition and its properties are inseparable.” Since the fusion protein comprising the Erwinia chrysanthemi L-asparaginase fused to the 201 alanine and proline polypeptide (PA#1(200) of SEQ ID NO:61) of Skerra has identical chemical structure as the fusion protein comprising L-asparaginase of SEQ ID NO:1 of the instant application fused to the 201 alanine and proline polypeptide PA#1(200) of SEQ ID NO:7 of the instant application, the recited property “wherein the fusion protein comprises greater asparaginase .. than the unmodified L-asparaginase” is necessarily present. MPEP 2112 II states that “[t]here is no requirement that a person of ordinary skill in the art would have recognized the inherent disclosure at the relevant time, but only that the subject matter is in fact inherent in the prior art reference.”.
Further, MPEP 2143.02. II. states that “[o]bviousness does not require absolute predictability, but at least some degree of predictability is required.”. In the instant case, one of ordinary skill in the art would have had a reasonable expectation of success at arriving the claimed fusion protein since Skerra discloses a fusion protein comprising of a random coil polypeptide consisting solely of prolines and alanines and biologically active polypeptides, such as an asparaginase, and Kotzia discloses an L-asparaginase.
Applicant argues that in view of the disclosure of Skerra, Kotzia, and P06608, one of ordinary skill in the art would not have bene motivated to make the claimed invention with a reasonable expectation of success to obtain a fusion protein that has “greater asparaginase or glutaminase activity that the L-asparaginase in an unmodified form”. Applicant argues that Skerra generally outlines a broad range of possible P/A polypeptide length about 50 to 3000 proline and alanine residues that may be used to increase the stability of other compounds. However, Applicant argues that Skerra does not provide guidance regarding which length of the P/A polypeptide are optimal to be used with what protein, let what length of the P/A polypeptide is optional for the activity of an L-asparaginase of SEQ ID NO:1. Applicant argues that the results of Skerra showed that the relevant filed of art is unpredictable because a protein has different secondary structure before or after it is fused with PA#1 (200) and the same coil PA#1(200 may have different effect on the secondary structure of different when forming fusion with these proteins. Applicant argues that Skerra provides no specific teaching on a fusion protein containing a L-asparaginase fused to a random coil polypeptide consisting solely of proline and alanine residues and having about 200 to about 400 amino acids residues in total.
This is not found persuasive.
Due to the amendment of claim 21, claims 21-24 are not directed to a fusion protein wherein the fusion protein has greater asparaginase or glutaminase activity that the L-asparaginase in an unmodified from. Therefore, Applicant’s arguments are moot for claims 21-24.
MPEP 2145. X.D.1. states that "the prior art's mere disclosure of more than one alternative does not constitute a teaching away from any of these alternatives because such disclosure does not criticize, discredit, or otherwise discourage the solution claimed.... MPEP 2123 states that "[d]isclosed examples and preferred embodiments do not constitute a teaching away from a broader disclosure or nonpreferred embodiments”. In the instant case, the results of Examples 10 and 15 of Skerra do not criticize, discredit, or otherwise discourage fusing the random coil PA#1(200) to biologically active polypeptides, such as asparaginase. Skerra teaches that random coil P/A polypeptides of about 200 alanine and prolines are capable of conferring increased in vivo/in vitro stability, which is useful for biologically active proteins comprising said random coil in medical applications (Column 8, line 59 through Column 9, line 28). Skerra discloses that the random coil P/A polypeptides does not interfere with the biological activity of the biologically active protein and is inert with respect to proteolysis in blood plasma and immunogenicity (Column 26, lines 25-50). MPEP 2143.02. II. states that “[o]bviousness does not require absolute predictability, but at least some degree of predictability is required.”. In the instant case, one of ordinary skill in the art would have had a reasonable expectation of success at arriving at the claimed fusion protein since Skerra discloses a fusion protein comprising of a random coil polypeptide consisting solely of prolines and alanines, such as the random coil polypeptide of PA#1(200) and biologically active polypeptides, such as an asparaginase, and Kotzia discloses an L-asparaginase having 100 sequence identity to SEQ ID NO:1.
Further, in response to applicant's argument that the cited references do not provide motivation to one having ordinary skill in the art to make the claimed fusion protein having greater asparaginase or glutaminase activity than the L- asparaginase in an unmodified form, the fact that the inventor has recognized another advantage which would flow naturally from following the suggestion of the prior art cannot be the basis for patentability when the differences would otherwise be obvious. See Ex parte Obiaya, 227 USPQ 58, 60 (Bd. Pat. App. & Inter. 1985). MPEP 2144. IV. states that the “reason or motivation to modify the reference may often suggest what the inventor has done, but for a different purpose or to solve a different problem. It is not necessary that the prior art suggest the combination to achieve the same advantage or result discovered by applicant.”. In the instant case, Skerra discloses specific examples of the random coil P/A polypeptides, such as PA#1(200) having the amino acid sequence of SEQ ID NO:61 (comprised in SEQ ID NO:27) consisting of 201 proline and alanine residues (Column 50, lines 43-54 and see the sequence alignment below). The PA#1(200) polypeptide of SEQ ID NO:61 of Skerra is identical to the PA#1(200) polypeptide of SEQ ID NO:7 of the instant application. Skerra teaches that random coil P/A polypeptides of about 200 alanine and prolines are capable of conferring increased in vivo/in vitro stability, which is useful for biologically active proteins comprising said random coil in medical applications (Column 8, line 59 through Column 9, line 28). Skerra discloses that the random coil P/A polypeptides does not interfere with the biological activity of the biologically active protein and is inert with respect to proteolysis in blood plasma and immunogenicity (Column 26, lines 25-50). Therefore, Skerra provides guidance on selecting different lengths of the random coils and their effect on the properties of the biologically active proteins.
Applicant argues that the claims are not obvious over the cited references because through innovative experimentation, were the inventors of the instant application able to arrive at the claimed invention. The inventors showed that, compared with the unmodified protein, the enzymatic activity of an L-asparaginase of SEQ ID NO:1 was not reduced, but was improved to 109% and 118% instead, when fused to P/A polypeptides comprising about 200 or about 400 proline and alanine amino acid residues. Such results are superior and unexpected over Skerra.
This is not found persuasive.
It appears that Applicant is referring to Example 5 of the instant application. Example 5 is limited to a fusion protein comprising PA #1(200) (SEQ ID NO:7)-Crisantaspase (L-asparaginase of SEQ ID NO:1) and a fusion protein comprising PA #1(400) (SEQ ID NO:9) -Crisantaspase (L-asparaginase of SEQ ID NO:1). Therefore, the results of PA #1(200)-Crisantaspase and PA #1(400)-Crisantaspase are not commensurate in scope with the claims (see MPEP 2145). The claims do not recite a fusion comprising the Erwinia L- asparaginase of SEQ ID NO:1 fused to SEQ ID NO: 7 or 9. Further, MPEP 2145.II. states that “[the] fact that appellant has recognized another advantage which would flow naturally from following the suggestion of the prior art cannot be the basis for patentability when the differences would otherwise be obvious.” In the instant case, the property “wherein the fusion protein comprises greater asparaginase .. than the unmodified L-asparaginase” recited in claim 1 is an inherent property of the fusion protein comprising the Erwinia chrysanthemi L-asparaginase fused to the 201 alanine and proline polypeptide (PA#1(200) of SEQ ID NO:61) of Skerra because said fusion protein has identical chemical structure as the fusion protein comprising L-asparaginase of SEQ ID NO:1 of the instant application fused to the 201 alanine and proline polypeptide PA#1(200) of SEQ ID NO:7 of the instant application as described in the specification of the instant application (Examples 5 and 8-9). The fusion protein described in Examples 5 and 8-9 has greater asparaginase activity than the unmodified L-asparaginase. MPEP 2112.01 states that “Products of identical chemical composition can not have mutually exclusive properties.” In re Spada, 911 F.2d 705, 709, 15 USPQ2d 1655, 1658 (Fed. Cir. 1990). A chemical composition and its properties are inseparable.” Since the fusion protein comprising the Erwinia chrysanthemi L-asparaginase fused to the 201 alanine and proline polypeptide (PA#1(200) of SEQ ID NO:61) of Skerra has identical chemical structure as the fusion protein comprising L-asparaginase of SEQ ID NO:1 of the instant application fused to the 201 alanine and proline polypeptide PA#1(200) of SEQ ID NO:7 of the instant application, the recited property “wherein the fusion protein comprises greater asparaginase .. than the unmodified L-asparaginase” is necessarily present and would flow naturally from following the suggestion of the prior art. MPEP 2112 II states that “[t]here is no requirement that a person of ordinary skill in the art would have recognized the inherent disclosure at the relevant time, but only that the subject matter is in fact inherent in the prior art reference.”
Hence the rejection is maintained.
Conclusion
Claims 1, 3-6, 8, and 19-30 are pending.
Claims 6, 20 and 25-30 are withdrawn.
Claims 1, 3-5, 8, 19, and 21-24 are rejected.
THIS ACTION IS MADE FINAL. Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to YONG D PAK whose telephone number is (571)272-0935. The examiner can normally be reached M-Th: 5:30 am - 3:30 pm.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Robert Mondesi can be reached on 408-918-7584. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/YONG D PAK/Primary Examiner, Art Unit 1652
Sequence alignment between SEQ ID NO:7 of the instant application (“Qy”) and SEQ ID NO:61 of Skerra (“Db”)
US-16-107-590-61
; Sequence 61, Application US/16107590
; Patent No. 10844094
; GENERAL INFORMATION
; APPLICANT: XL-protein GmbH
; APPLICANT:Technische UniversitAt MUnchen
; APPLICANT:SKERRA, Arne
; APPLICANT:BINDER, Uli
; APPLICANT:SCHLAPSCHY, Martin
; TITLE OF INVENTION: Biosynthetic proline/alanine random coil polypeptides and their
; TITLE OF INVENTION:uses
; FILE REFERENCE: S1467 PCT S3
; CURRENT APPLICATION NUMBER: US/16/107,590
; CURRENT FILING DATE: 2018-08-21
; PRIOR APPLICATION NUMBER: US 14/939,626
; PRIOR FILING DATE: 2016-01-20
; NUMBER OF SEQ ID NOS: 61
; SOFTWARE: PatentIn version 3.3
; SEQ ID NO 61
; LENGTH: 201
; TYPE: PRT
; ORGANISM: artificial sequence
; FEATURE:
; NAME/KEY: source
; OTHER INFORMATION: /note="Description of artificial sequence: PA#1(200)
; OTHER INFORMATION:polypeptide/polymer used for the preparation of drug conjugates
; OTHER INFORMATION:(made by ligation of 10 20mer encoding gene cassettes, including
; OTHER INFORMATION:one additional C-terminal Ala residue resulting from the downstream
; OTHER INFORMATION:ligation site)"
US-16-107-590-61
Query Match 100.0%; Score 1014; DB 3; Length 201;
Best Local Similarity 100.0%;
Matches 201; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 60
Qy 61 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 120
Qy 121 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 180
Qy 181 AAPAAPAPAAPAAPAPAAPAA 201
|||||||||||||||||||||
Db 181 AAPAAPAPAAPAAPAPAAPAA 201
Sequence alignment between SEQ ID NO:7 of the instant application (“Qy”) and SEQ ID NO:27 of Skerra (“Db”)
US-16-107-590-27
; Sequence 27, Application US/16107590
; Patent No. 10844094
; GENERAL INFORMATION
; APPLICANT: XL-protein GmbH
; APPLICANT:Technische UniversitAt MUnchen
; APPLICANT:SKERRA, Arne
; APPLICANT:BINDER, Uli
; APPLICANT:SCHLAPSCHY, Martin
; TITLE OF INVENTION: Biosynthetic proline/alanine random coil polypeptides and their
; TITLE OF INVENTION:uses
; FILE REFERENCE: S1467 PCT S3
; CURRENT APPLICATION NUMBER: US/16/107,590
; CURRENT FILING DATE: 2018-08-21
; PRIOR APPLICATION NUMBER: US 14/939,626
; PRIOR FILING DATE: 2016-01-20
; NUMBER OF SEQ ID NOS: 61
; SOFTWARE: PatentIn version 3.3
; SEQ ID NO 27
; LENGTH: 417
; TYPE: PRT
; ORGANISM: artificial sequence
; FEATURE:
; NAME/KEY: source
; OTHER INFORMATION: /note="Description of artificial sequence: amino acid sequence of
; OTHER INFORMATION:the Fab light chain fused with the PA#1(200) polymer as encoded
; OTHER INFORMATION:on pFab-PA#1(200)"
US-16-107-590-27
Query Match 100.0%; Score 1014; DB 3; Length 417;
Best Local Similarity 100.0%;
Matches 201; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 217 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 276
Qy 61 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 277 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 336
Qy 121 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 337 AAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPAAAPAAPAPAAPAAPAPAAPA 396
Qy 181 AAPAAPAPAAPAAPAPAAPAA 201
|||||||||||||||||||||
Db 397 AAPAAPAPAAPAAPAPAAPAA 417
Sequence alignment between SEQ ID NO:5 of the instant application (“Qy”) and SEQ ID NO:61 of Skerra (“Db”)
US-13-697-569-61
Sequence 61, US/13697569
Patent No. 9221882
GENERAL INFORMATION
APPLICANT: XL-protein GmbH
APPLICANT: Technische UniversitAt MUnchen
APPLICANT: SKERRA, Arne
APPLICANT: BINDER, Uli
APPLICANT: SCHLAPSCHY, Martin
TITLE OF INVENTION: Biosynthetic proline/alanine random coil polypeptides and their
TITLE OF INVENTION: uses
FILE REFERENCE: S1467 PCT S3
CURRENT APPLICATION NUMBER: US/13/697,569
CURRENT FILING DATE: 2012-11-13
NUMBER OF SEQ ID NOS: 61
SEQ ID NO 61
LENGTH: 201
TYPE: PRT
ORGANISM: artificial sequence
FEATURE:
NAME/KEY: source
OTHER INFORMATION: /note="Description of artificial sequence: PA#1(200)
polypeptide/polymer used for the preparation of drug conjugates
(made by ligation of 10 20mer encoding gene cassettes, including
one additional C-terminal Ala residue resulting from the downstream
ligation site)"
Query Match 100.0%; Score 101; Length 201;
Best Local Similarity 100.0%;
Matches 20; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AAPAAPAPAAPAAPAPAAPA 20
||||||||||||||||||||
Db 1 AAPAAPAPAAPAAPAPAAPA 20
Sequence alignment between SEQ ID NO:1 (“Qy”) and the L-asparaginase of Kotzia (“Db”)
ASPG_DICCH
ID ASPG_DICCH Reviewed; 348 AA.
AC P06608;
DT 01-JAN-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1988, sequence version 1.
DT 25-MAY-2022, entry version 110.
DE RecName: Full=L-asparaginase;
DE Short=L-ASNase;
DE EC=3.5.1.1;
DE AltName: Full=L-asparagine amidohydrolase;
DE Flags: Precursor;
GN Name=ansB; Synonyms=asn;
OS Dickeya chrysanthemi (Pectobacterium chrysanthemi) (Erwinia chrysanthemi).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Pectobacteriaceae; Dickeya.
OX NCBI_TaxID=556;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NCPPB 1066;
RX PubMed=3026924; DOI=10.1016/0378-1119(86)90163-0;
RA Minton N.P., Bullman H.M.S., Scawen M.D., Atkinson T., Gilbert H.J.;
RT "Nucleotide sequence of the Erwinia chrysanthemi NCPPB 1066 L-asparaginase
RT gene.";
RL Gene 46:25-35(1986).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NCPPB 1125;
RX PubMed=3194219; DOI=10.1093/nar/16.21.10385;
RA Filpula D., Nagle J.W., Pulford S., Anderson D.M.;
RT "Sequence of L-asparaginase gene from Erwinia chrysanthemi NCPPB 1125.";
RL Nucleic Acids Res. 16:10385-10385(1988).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS).
RX PubMed=3379033; DOI=10.1016/s0021-9258(18)68344-9;
RA Tanaka S., Robinson E.A., Appella E., Miller M., Ammon H.L., Roberts J.,
RA Weber I.T., Wlodawer A.;
RT "Structures of amidohydrolases. Amino acid sequence of a glutaminase-
RT asparaginase from Acinetobacter glutaminasificans and preliminary
RT crystallographic data for an asparaginase from Erwinia chrysanthemi.";
RL J. Biol. Chem. 263:8583-8591(1988).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) IN COMPLEX WITH ASPARTIC ACID, ACTIVE
RP SITE, AND SUBUNIT.
RX PubMed=8348975; DOI=10.1016/0014-5793(93)80943-o;
RA Miller M.M., Rao J.K.M., Wlodawer A., Gribskov M.R.;
RT "A left-handed crossover involved in amidohydrolase catalysis. Crystal
RT structure of Erwinia chrysanthemi L-asparaginase with bound L-aspartate.";
RL FEBS Lett. 328:275-279(1993).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) OF 22-348 IN COMPLEXES WITH GLUTAMIC
RP ACID; ASPARTIC ACID AND SUCCINIC ACID.
RX PubMed=11341830; DOI=10.1021/bi0029595;
RA Aghaiypour K., Wlodawer A., Lubkowski J.;
RT "Structural basis for the activity and substrate specificity of Erwinia
RT chrysanthemi L-asparaginase.";
RL Biochemistry 40:5655-5664(2001).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 22-348 IN COMPLEX WITH INHIBITOR
RP 6-DIAZO-5-OXY-NORLEUCINE, AND ACTIVE SITE.
RX PubMed=11755201; DOI=10.1016/s0167-4838(01)00270-9;
RA Aghaiypour K., Wlodawer A., Lubkowski J.;
RT "Do bacterial L-asparaginases utilize a catalytic triad Thr-Tyr-Glu?";
RL Biochim. Biophys. Acta 1550:117-128(2001).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.0 ANGSTROMS) OF 22-348, AND SUBUNIT.
RX PubMed=12499544; DOI=10.1107/s0907444902019443;
RA Lubkowski J., Dauter M., Aghaiypour K., Wlodawer A., Dauter Z.;
RT "Atomic resolution structure of Erwinia chrysanthemi L-asparaginase.";
RL Acta Crystallogr. D 59:84-92(2003).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-asparagine = L-aspartate + NH4(+);
CC Xref=Rhea:RHEA:21016, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:29991, ChEBI:CHEBI:58048; EC=3.5.1.1;
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:11755201,
CC ECO:0000269|PubMed:12499544, ECO:0000269|PubMed:8348975}.
CC -!- PHARMACEUTICAL: Available under the name Erwinase (Beaufour Ipsen).
CC Used as an antineoplastic in chemotherapy. Reduces the quantity of
CC asparagine available to cancer cells.
CC -!- MISCELLANEOUS: The sequence of strain NCPPB 1066 is shown.
CC -!- SIMILARITY: Belongs to the asparaginase 1 family. {ECO:0000305}.
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DR EMBL; X14777; CAA32884.1; -; Genomic_DNA.
DR EMBL; X12746; CAA31239.1; -; Genomic_DNA.
DR PIR; A26054; A26054.
DR PDB; 1HFJ; X-ray; 2.40 A; A/C=22-348.
DR PDB; 1HFK; X-ray; 2.17 A; A/C=22-348.
DR PDB; 1HFW; X-ray; 1.80 A; A/B/C/D=22-348.
DR PDB; 1HG0; X-ray; 1.90 A; A/B/C/D=22-348.
DR PDB; 1HG1; X-ray; 1.80 A; A/B/C/D=22-348.
DR PDB; 1JSL; X-ray; 1.70 A; A/B/C/D=22-348.
DR PDB; 1JSR; X-ray; 1.70 A; A/B/C/D=22-348.
DR PDB; 1O7J; X-ray; 1.00 A; A/B/C/D=22-348.
DR PDB; 5F52; X-ray; 1.63 A; A/B/C/D=23-348.
DR PDB; 5HW0; X-ray; 1.70 A; A/B/C/D=23-348.
DR PDB; 5I3Z; X-ray; 2.05 A; A/B/C/D=23-348.
DR PDB; 5I48; X-ray; 1.50 A; A/B/C/D=23-348.
DR PDB; 5I4B; X-ray; 1.60 A; A/B/C=23-348.
DR PDB; 6PAE; X-ray; 1.60 A; A/B/C/D=22-348.
DR PDBsum; 1HFJ; -.
DR PDBsum; 1HFK; -.
DR PDBsum; 1HFW; -.
DR PDBsum; 1HG0; -.
DR PDBsum; 1HG1; -.
DR PDBsum; 1JSL; -.
DR PDBsum; 1JSR; -.
DR PDBsum; 1O7J; -.
DR PDBsum; 5F52; -.
DR PDBsum; 5HW0; -.
DR PDBsum; 5I3Z; -.
DR PDBsum; 5I48; -.
DR PDBsum; 5I4B; -.
DR PDBsum; 6PAE; -.
DR AlphaFoldDB; P06608; -.
DR SMR; P06608; -.
DR ChEMBL; CHEMBL1075190; -.
DR DrugBank; DB02233; 6-hydroxy-D-norleucine.
DR DrugBank; DB03412; 6-hydroxynorleucine.
DR DrugBank; DB02655; D-Aspartic Acid.
DR Allergome; 8366; Erw ch Asparaginase.
DR PRIDE; P06608; -.
DR BRENDA; 3.5.1.1; 2141.
DR EvolutionaryTrace; P06608; -.
DR GO; GO:0004067; F:asparaginase activity; IEA:UniProtKB-EC.
DR GO; GO:0006528; P:asparagine metabolic process; IEA:InterPro.
DR Gene3D; 3.40.50.1170; -; 1.
DR Gene3D; 3.40.50.40; -; 1.
DR InterPro; IPR004550; AsnASE_II.
DR InterPro; IPR036152; Asp/glu_Ase-like_sf.
DR InterPro; IPR006034; Asparaginase/glutaminase-like.
DR InterPro; IPR020827; Asparaginase/glutaminase_AS1.
DR InterPro; IPR027475; Asparaginase/glutaminase_AS2.
DR InterPro; IPR040919; Asparaginase_C.
DR InterPro; IPR027473; L-asparaginase_C.
DR InterPro; IPR027474; L-asparaginase_N.
DR InterPro; IPR037152; L-asparaginase_N_sf.
DR Pfam; PF00710; Asparaginase; 1.
DR Pfam; PF17763; Asparaginase_C; 1.
DR PIRSF; PIRSF001220; L-ASNase_gatD; 1.
DR PRINTS; PR00139; ASNGLNASE.
DR SMART; SM00870; Asparaginase; 1.
DR SUPFAM; SSF53774; SSF53774; 1.
DR TIGRFAMs; TIGR00520; asnASE_II; 1.
DR PROSITE; PS00144; ASN_GLN_ASE_1; 1.
DR PROSITE; PS00917; ASN_GLN_ASE_2; 1.
DR PROSITE; PS51732; ASN_GLN_ASE_3; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Pharmaceutical; Signal.
FT SIGNAL 1..21
FT CHAIN 22..348
FT /note="L-asparaginase"
FT /id="PRO_0000002358"
FT DOMAIN 26..348
FT /note="Asparaginase/glutaminase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01068"
FT REGION 116..117
FT /note="Substrate binding"
FT ACT_SITE 36
FT /note="O-isoaspartyl threonine intermediate"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10099,
FT ECO:0000255|PROSITE-ProRule:PRU10100,
FT ECO:0000269|PubMed:11755201, ECO:0000269|PubMed:8348975"
FT BINDING 83
FT /note="Substrate"
FT VARIANT 177
FT /note="L -> I (in strain: NCPPB 1125)"
FT VARIANT 199
FT /note="K -> R (in strain: NCPPB 1125)"
FT VARIANT 288
FT /note="M -> L (in strain: NCPPB 1125)"
FT VARIANT 295
FT /note="I -> M (in strain: NCPPB 1125)"
FT STRAND 27..35
FT /evidence="ECO:0007829|PDB:1O7J"
FT HELIX 36..38
FT /evidence="ECO:0007829|PDB:1O7J"
FT STRAND 42..45
FT /evidence="ECO:0007829|PDB:5HW0"
FT STRAND 48..50
FT /evidence="ECO:0007829|PDB:5F52"
FT STRAND 51..54
FT /evidence="ECO:0007829|PDB:5I48"
FT HELIX 57..63
FT /evidence="ECO:0007829|PDB:1O7J"
FT HELIX 65..69
FT /evidence="ECO:0007829|PDB:1O7J"
FT STRAND 72..81
FT /evidence="ECO:0007829|PDB:1O7J"
FT HELIX 83..85
FT /evidence="ECO:0007829|PDB:1O7J"
FT HELIX 88..102
FT /evidence="ECO:0007829|PDB:1O7J"
FT STRAND 109..113
FT /evidence="ECO:0007829|PDB:1O7J"
FT TURN 116..118
FT /evidence="ECO:0007829|PDB:5I48"
FT HELIX 119..129
FT /evidence="ECO:0007829|PDB:1O7J"
FT STRAND 136..139
Query Match 100.0%; Score 1644; DB 1; Length 348;
Best Local Similarity 100.0%;
Matches 327; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ADKLPNIVILATGGTIAGSAATGTQTTGYKAGALGVDTLINAVPEVKKLANVKGEQFSNM 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 22 ADKLPNIVILATGGTIAGSAATGTQTTGYKAGALGVDTLINAVPEVKKLANVKGEQFSNM 81
Qy 61 ASENMTGDVVLKLSQRVNELLARDDVDGVVITHGTDTVEESAYFLHLTVKSDKPVVFVAA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 82 ASENMTGDVVLKLSQRVNELLARDDVDGVVITHGTDTVEESAYFLHLTVKSDKPVVFVAA 141
Qy 121 MRPATAISADGPMNLLEAVRVAGDKQSRGRGVMVVLNDRIGSARYITKTNASTLDTFKAN 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 142 MRPATAISADGPMNLLEAVRVAGDKQSRGRGVMVVLNDRIGSARYITKTNASTLDTFKAN 201
Qy 181 EEGYLGVIIGNRIYYQNRIDKLHTTRSVFDVRGLTSLPKVDILYGYQDDPEYLYDAAIQH 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 202 EEGYLGVIIGNRIYYQNRIDKLHTTRSVFDVRGLTSLPKVDILYGYQDDPEYLYDAAIQH 261
Qy 241 GVKGIVYAGMGAGSVSVRGIAGMRKAMEKGVVVIRSTRTGNGIVPPDEELPGLVSDSLNP 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 262 GVKGIVYAGMGAGSVSVRGIAGMRKAMEKGVVVIRSTRTGNGIVPPDEELPGLVSDSLNP 321
Qy 301 AHARILLMLALTRTSDPKVIQEYFHTY 327
|||||||||||||||||||||||||||
Db 322 AHARILLMLALTRTSDPKVIQEYFHTY 348