Reagent to label proteins via lysine isopeptide bonds
Abstract
Covalently cross-linked pilus polymers displayed on the cell surface of Gram-positive bacteria are assembled by class C sortase enzymes. These pilus-specific transpeptidases located on the bacterial membrane catalyze a two-step protein ligation reaction—first, cleaving the LPXTG motif of one pilin protomer to form an acyl-enzyme intermediate, and second, joining the terminal threonine to the nucleophilic lysine residue residing within the pilin motif of another pilin protomer. Informed by the high-resolution crystal structures of corynebacterial pilus-specific sortase (SrtA) and by developing structural variants of the sortase enzyme whose catalytic pocket has been unmasked by activating mutations, we have developed new reagents capable of forming isopeptide bonds in vitro. The reagents disclosed herein can catalyze ligation of isolated SpaA domains in vitro provide a facile and versatile new platform for protein engineering and bio-conjugation that has major implications for biotechnology.
- Inventors:
- Issue Date:
- Research Org.:
- Univ. of California, Los Angeles, CA (United States); Univ. of Texas, Austin, TX (United States)
- Sponsoring Org.:
- USDOE; National Institutes of Health (NIH)
- OSTI Identifier:
- 1998327
- Patent Number(s):
- 11634699
- Application Number:
- 17/052,116
- Assignee:
- The Regents of the University of California (Oakland, CA); The Board of Regents of The University of Texas System (Austin, TX)
- DOE Contract Number:
- FC02-02ER63421; DE017382; DE025015
- Resource Type:
- Patent
- Resource Relation:
- Patent File Date: 05/01/2019
- Country of Publication:
- United States
- Language:
- English
Citation Formats
Clubb, Robert T., Amer, Brendan Rayhan, Fu, Janine Y., McConnell, Scott, Ton-That, Hung, and Chang, Chungyu. Reagent to label proteins via lysine isopeptide bonds. United States: N. p., 2023.
Web.
Clubb, Robert T., Amer, Brendan Rayhan, Fu, Janine Y., McConnell, Scott, Ton-That, Hung, & Chang, Chungyu. Reagent to label proteins via lysine isopeptide bonds. United States.
Clubb, Robert T., Amer, Brendan Rayhan, Fu, Janine Y., McConnell, Scott, Ton-That, Hung, and Chang, Chungyu. Tue .
"Reagent to label proteins via lysine isopeptide bonds". United States. https://www.osti.gov/servlets/purl/1998327.
@article{osti_1998327,
title = {Reagent to label proteins via lysine isopeptide bonds},
author = {Clubb, Robert T. and Amer, Brendan Rayhan and Fu, Janine Y. and McConnell, Scott and Ton-That, Hung and Chang, Chungyu},
abstractNote = {Covalently cross-linked pilus polymers displayed on the cell surface of Gram-positive bacteria are assembled by class C sortase enzymes. These pilus-specific transpeptidases located on the bacterial membrane catalyze a two-step protein ligation reaction—first, cleaving the LPXTG motif of one pilin protomer to form an acyl-enzyme intermediate, and second, joining the terminal threonine to the nucleophilic lysine residue residing within the pilin motif of another pilin protomer. Informed by the high-resolution crystal structures of corynebacterial pilus-specific sortase (SrtA) and by developing structural variants of the sortase enzyme whose catalytic pocket has been unmasked by activating mutations, we have developed new reagents capable of forming isopeptide bonds in vitro. The reagents disclosed herein can catalyze ligation of isolated SpaA domains in vitro provide a facile and versatile new platform for protein engineering and bio-conjugation that has major implications for biotechnology.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2023},
month = {4}
}
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