Reversibly Reactive Affinity Selection-Mass Spectrometry Enables Identification of Covalent Peptide Binders.

Autor:	Zhang P; Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States., Ye X; Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States., Wang JCK; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Baddock HT; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Jensvold Z; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Foe IT; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Loas A; Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States., Eaton DL; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Hao Q; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Nile AH; Calico Life Sciences LLC, 1170 Veterans Boulevard, South San Francisco, California 94080, United States., Pentelute BL; Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.; The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 500 Main Street, Cambridge, Massachusetts 02142, United States.; Center for Environmental Health Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.; Broad Institute of MIT and Harvard, 415 Main Street, Cambridge, Massachusetts 02142, United States.
Jazyk:	angličtina
Zdroj:	Journal of the American Chemical Society [J Am Chem Soc] 2024 Jun 05; Vol. 146 (22), pp. 15627-15639. Date of Electronic Publication: 2024 May 21.
DOI:	10.1021/jacs.4c05571
Abstrakt:	Covalent peptide binders have found applications as activity-based probes and as irreversible therapeutic inhibitors. Currently, there is no rapid, label-free, and tunable affinity selection platform to enrich covalent reactive peptide binders from synthetic libraries. We address this challenge by developing a reversibly reactive affinity selection platform termed ReAct-ASMS enabled by tandem high-resolution mass spectrometry (MS/MS) to identify covalent peptide binders to native protein targets. It uses mixed disulfide-containing peptides to build reversible peptide-protein conjugates that can enrich for covalent variants, which can be sequenced by MS/MS after reduction. Using this platform, we identified covalent peptide binders against two oncoproteins, human papillomavirus 16 early protein 6 (HPV16 E6) and peptidyl-prolyl cis - trans isomerase NIMA-interacting 1 protein (Pin1). The resulting peptide binders efficiently and selectively cross-link Cys58 of E6 at 37 °C and Cys113 of Pin1 at room temperature, respectively. ReAct-ASMS enables the identification of highly selective covalent peptide binders for diverse molecular targets, introducing an applicable platform to assist preclinical therapeutic development pipelines.
Databáze:	MEDLINE
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