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Chemoproteomic Strategy to Quantitatively Monitor Transnitrosation Uncovers Functionally Relevant S-Nitrosation Sites on Cathepsin D and HADH2. Cell Chem Biol 2016 Jun 23;23(6):727-37

Date

06/14/2016

Pubmed ID

27291402

Pubmed Central ID

PMC4920707

DOI

10.1016/j.chembiol.2016.05.008

Scopus ID

2-s2.0-84976439988 (requires institutional sign-in at Scopus site)   39 Citations

Abstract

S-Nitrosoglutathione (GSNO) is an endogenous transnitrosation donor involved in S-nitrosation of a variety of cellular proteins, thereby regulating diverse protein functions. Quantitative proteomic methods are necessary to establish which cysteine residues are most sensitive to GSNO-mediated transnitrosation. Here, a competitive cysteine-reactivity profiling strategy was implemented to quantitatively measure the sensitivity of >600 cysteine residues to transnitrosation by GSNO. This platform identified a subset of cysteine residues with a high propensity for GSNO-mediated transnitrosation. Functional characterization of previously unannotated S-nitrosation sites revealed that S-nitrosation of a cysteine residue distal to the 3-hydroxyacyl-CoA dehydrogenase type 2 (HADH2) active site impaired catalytic activity. Similarly, S-nitrosation of a non-catalytic cysteine residue in the lysosomal aspartyl protease cathepsin D (CTSD) inhibited proteolytic activation. Together, these studies revealed two previously uncharacterized cysteine residues that regulate protein function, and established a chemical-proteomic platform with capabilities to determine substrate specificity of other cellular transnitrosation agents.

Author List

Zhou Y, Wynia-Smith SL, Couvertier SM, Kalous KS, Marletta MA, Smith BC, Weerapana E

Authors

Brian C. Smith PhD Associate Professor in the Biochemistry department at Medical College of Wisconsin
Sarah L. Wynia Smith Research Scientist II in the Biochemistry department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

3-Hydroxyacyl CoA Dehydrogenases
Cathepsin D
Humans
MCF-7 Cells
Nitrosation
Proteomics