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WO2015127365A3 - Calcium-independent sortase a mutants - Google Patents

Calcium-independent sortase a mutants Download PDF

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Publication number
WO2015127365A3
WO2015127365A3 PCT/US2015/017116 US2015017116W WO2015127365A3 WO 2015127365 A3 WO2015127365 A3 WO 2015127365A3 US 2015017116 W US2015017116 W US 2015017116W WO 2015127365 A3 WO2015127365 A3 WO 2015127365A3
Authority
WO
WIPO (PCT)
Prior art keywords
residue
sortase
calcium
amino acid
mutants
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2015/017116
Other languages
French (fr)
Other versions
WO2015127365A2 (en
Inventor
Jessica Ingram
Hidde Ploegh
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Whitehead Institute for Biomedical Research
Original Assignee
Whitehead Institute for Biomedical Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Whitehead Institute for Biomedical Research filed Critical Whitehead Institute for Biomedical Research
Publication of WO2015127365A2 publication Critical patent/WO2015127365A2/en
Publication of WO2015127365A3 publication Critical patent/WO2015127365A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/22—Cysteine endopeptidases (3.4.22)
    • C12Y304/2207—Sortase A (3.4.22.70)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Disclosed herein are calcium-independent sortase A mutants exhibiting increased catalytic activity compared to wild-type sortase A. In some aspects, the disclosure provides a sortase A mutant comprising at least three amino acid substitutions relative to a wild-type sortase A, wherein the amino acid substitutions comprise a) a K residue at position 105; b) a Q or A residue at position 108; and c) at least one amino acid substitution selected from the group consisting of i) a R residue at position 94; ii) a S residue at position 94; iii) a N residue at position.160; iv) a A residue at position 165; v) a E residue at 5 position 190; and vi) a T residue at position 196. In some embodiments, the mutant comprises a deletion of amino acids 2-25.
PCT/US2015/017116 2014-02-21 2015-02-23 Calcium-independent sortase a mutants Ceased WO2015127365A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201461943042P 2014-02-21 2014-02-21
US61/943,042 2014-02-21

Publications (2)

Publication Number Publication Date
WO2015127365A2 WO2015127365A2 (en) 2015-08-27
WO2015127365A3 true WO2015127365A3 (en) 2015-10-15

Family

ID=53879254

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2015/017116 Ceased WO2015127365A2 (en) 2014-02-21 2015-02-23 Calcium-independent sortase a mutants

Country Status (1)

Country Link
WO (1) WO2015127365A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018005716A2 (en) * 2016-07-01 2018-01-04 Denali Therapeutics Inc. Albumin variants for enhanced serum half-life
CN116917484A (en) * 2021-01-28 2023-10-20 启德医药科技（苏州）有限公司 Ligase fusion proteins and their applications

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6773706B2 (en) * 1999-04-15 2004-08-10 The Regents Of The University Of California Identification of sortase gene
WO2013016653A1 (en) * 2011-07-28 2013-01-31 Cell Signaling Technology, Inc. Multi component detection

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6773706B2 (en) * 1999-04-15 2004-08-10 The Regents Of The University Of California Identification of sortase gene
WO2013016653A1 (en) * 2011-07-28 2013-01-31 Cell Signaling Technology, Inc. Multi component detection

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE NCBI, Genbank 17 November 2009 (2009-11-17), "Staphylococcus aureus subsp. aureus ED98", XP055231327, Database accession no. ACY12423 *
HIRAKAWA ET AL.: "Design of Ca2+-independent Staphylococcus aureus sortase A mutants", BIOTECHNOLOGY AND BIOENGINEERING, vol. 109, no. 12, 4 July 2012 (2012-07-04), pages 2955 - 2961, XP055210375 *
LOWDER ET AL.: "Recent human-to-poultry host jump, adaptation, and pandemic spread of Staphylococcus aureus", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 106, no. 46, 17 November 2009 (2009-11-17), pages 19545 - 19550, XP055039038 *
MARESSO ET AL.: "Sortase as a Target of Anti-Infective Therapy", PHARMACOLOGICAL REVIEWS, vol. 60, no. 1, 2008, pages 128 - 141, XP055231326 *

Also Published As

Publication number Publication date
WO2015127365A2 (en) 2015-08-27

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