Development of an α-Klotho Recognizing High-Affinity Peptide Probe from In-Solution Enrichment.

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., Smith CL; AbbVie Bioresearch Center, 100 Research Drive, Worcester, Massachusetts 01605, United States., Sousa S; AbbVie Bioresearch Center, 100 Research Drive, Worcester, Massachusetts 01605, 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., Preciado López M; 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:	JACS Au [JACS Au] 2024 Apr 10; Vol. 4 (4), pp. 1334-1344. Date of Electronic Publication: 2024 Apr 10 (Print Publication: 2024).
DOI:	10.1021/jacsau.3c00650
Abstrakt:	The kidney, parathyroid gland, and choroid plexus express the aging-related transmembrane protein α-Klotho, a coreceptor of the fibroblast growth factor 23 (FGF23) receptor complex. Reduced α-Klotho levels are correlated with chronic kidney disease and other age-related diseases, wherein they are released from membranes into circulation. Klotho's potential physiological action as a hormone is of current scientific interest. Part of the challenges associated with advancing these studies, however, has been the long-standing difficulty in detecting soluble α-Klotho in biofluids. Here, we describe the discovery of peptides that recognize α-Klotho with high affinity and selectivity by applying in-solution size-exclusion-based affinity selection-mass spectrometry (AS-MS). After two rounds of AS-MS and subsequent N-terminal modifications, the peptides improved their binding affinity to α-Klotho by approximately 2300-fold compared to the reported starting peptide Pep-10, previously designed based on the C-terminal region of FGF23. The lead peptide binders were shown to enrich α-Klotho from cell lysates and to label α-Klotho in kidney cells. Our results further support the utility of in-solution, label-free AS-MS protocols to discover peptide-based binders to target proteins of interest with high affinity and selectivity, resulting in functional probes for biological studies. Competing Interests: The authors declare the following competing financial interest(s): B.L.P. is a co-founder and/or member of the scientific advisory board of several companies focusing on the development of protein and peptide therapeutics. (© 2024 The Authors. Published by American Chemical Society.)
Databáze:	MEDLINE
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