Open AccessEngineering of a Small Protein Scaffold To Recognize Sulfotyrosine with High Specificity
Author(s) -
Justin Lawrie,
Sean Waldrop,
Anya Morozov,
Wei Niu,
Jiantao Guo
Publication year - 2021
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.1c00382
Subject(s) - sh2 domain , computational biology , mutant , proto oncogene tyrosine protein kinase src , protein engineering , biochemistry , biology , protein–protein interaction , tyrosine , microbiology and biotechnology , chemistry , signal transduction , enzyme , gene
Protein tyrosine O -sulfation is an essential post-translational modification required for effective biological processes such as hemostasis, inflammatory response, and visual phototransduction. Because of its unstable nature under mass spectrometry conditions and residing on low-abundance cell surface proteins, sulfated tyrosine (sulfotyrosine) residues are difficult to detect or analyze. Enrichment of sulfotyrosine-containing proteins (sulfoproteins) from complex biological samples are typically required before analysis. In this work, we seek to engineer the phosphotyrosine binding pocket of a Src Homology 2 (SH2) domain to act as an antisulfotyrosine antibody mimic. Using tailored selection schemes, several SH2 mutants are identified with high affinity and specificity to sulfotyrosine. Further molecular docking simulations highlight potential mechanisms supporting observed characteristics of these SH2 mutants. Utilities of the evolved SH2 mutants were demonstrated by the detection and enrichment of sulfoproteins.