Chemoproteomic Profiling of Phosphoaspartate Modifications in Prokaryotes.

Autor: Chang JW; Department of Chemistry, Institute for Genomics and Systems Biology, The University of Chicago, 929 E. 57th Street, Chicago, IL, 60637, USA., Montgomery JE; Department of Chemistry, Institute for Genomics and Systems Biology, The University of Chicago, 929 E. 57th Street, Chicago, IL, 60637, USA., Lee G; Department of Chemistry, Institute for Genomics and Systems Biology, The University of Chicago, 929 E. 57th Street, Chicago, IL, 60637, USA., Moellering RE; Department of Chemistry, Institute for Genomics and Systems Biology, The University of Chicago, 929 E. 57th Street, Chicago, IL, 60637, USA.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2018 Nov 26; Vol. 57 (48), pp. 15712-15716. Date of Electronic Publication: 2018 Nov 07.
DOI: 10.1002/anie.201809059
Abstrakt: Phosphorylation at aspartic acid residues represents an abundant and critical post-translational modification (PTM) in prokaryotes. In contrast to most characterized PTMs, such as phosphorylation at serine or threonine, the phosphoaspartate moiety is intrinsically labile, and therefore incompatible with common proteomic profiling methods. Herein, we report a nucleophilic, desthiobiotin-containing hydroxylamine (DBHA) chemical probe that covalently labels modified aspartic acid residues in native proteomes. DBHA treatment coupled with LC-MS/MS analysis enabled detection of known phosphoaspartate modifications, as well as novel aspartic acid sites in the E. coli proteome. Coupled with isotopic labelling, DBHA-dependent proteomic profiling also permitted global quantification of changes in endogenous protein modification status, as demonstrated with the detection of increased E. coli OmpR phosphorylation, but not abundance, in response to changes in osmolarity.
(© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
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