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PMID: 21085121 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Quantitative reactivity profiling predicts functional cysteines in proteomes.

Nature ·Vol. 468 ·No. 7325 ·2010-12-09 ·Pages 790-5

Weerapana E, Wang C, Simon GM, Richter F, Khare S, Dillon MB, Bachovchin DA, Mowen K, Baker D, Cravatt BF

Abstract

Cysteine is the most intrinsically nucleophilic amino acid in proteins, where its reactivity is tuned to perform diverse biochemical functions. The absence of a consensus sequence that defines functional cysteines in proteins has hindered their discovery and characterization. Here we describe a proteomics method to profile quantitatively the intrinsic reactivity of cysteine residues en masse directly in native biological systems. Hyper-reactivity was a rare feature among cysteines and it was found to specify a wide range of activities, including nucleophilic and reductive catalysis and sites of oxidative modification. Hyper-reactive cysteines were identified in several proteins of uncharacterized function, including a residue conserved across eukaryotic phylogeny that we show is required for yeast viability and is involved in iron-sulphur protein biogenesis. We also demonstrate that quantitative reactivity profiling can form the basis for screening and functional assignment of cysteines in computationally designed proteins, where it discriminated catalytically active from inactive cysteine hydrolase designs.

MeSH Terms
Animals Biocatalysis Cell Line, Tumor Conserved Sequence Cysteine/analysis,metabolism Humans Hydrolases/chemistry,metabolism Iron-Sulfur Proteins/biosynthesis Liver/metabolism Mice Myocardium/metabolism Nuclear Proteins/chemistry,metabolism Oxidation-Reduction Protein Engineering Protein Hydrolysates Protein-Arginine N-Methyltransferases/chemistry,metabolism Proteins/chemistry,metabolism Proteome/chemistry,metabolism Proteomics/methods Repressor Proteins/chemistry,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/chemistry,genetics,metabolism
Chemicals
FAM9B protein, human Iron-Sulfur Proteins Nuclear Proteins Protein Hydrolysates Proteins Proteome Repressor Proteins Saccharomyces cerevisiae Proteins YHR122W protein, S cerevisiae hydrolysin PRMT1 protein, human Protein-Arginine N-Methyltransferases Hydrolases Cysteine
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Weerapana Eranthie
The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Wang Chu
Simon Gabriel M
Richter Florian
Khare Sagar
Dillon Myles B D
Bachovchin Daniel A
Mowen Kerri
Baker David
Cravatt Benjamin F
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Article Info
Journal
Nature
Abbr.
Nature
ISSN
1476-4687
DOI
10.1038/nature09472
Published
2010-12-09
Epub
2010-00-17
Pages
790-5
Language
English
Region
England
NLM ID
0410462
PMCID
PMC3058684
Subset
IM
Grants
NCI NIH HHS · CA087660 · United States
NCI NIH HHS · R37 CA087660 · United States
NIMH NIH HHS · U54 MH084512 · United States
NIGMS NIH HHS · R01 GM085117 · United States
NIGMS NIH HHS · R01 GM090294 · United States
NCI NIH HHS · R01 CA087660-09 · United States
NCI NIH HHS · R37 CA087660-10 · United States
NIMH NIH HHS · MH084512 · United States
NIMH NIH HHS · U54 MH084512-030004 · United States
NCI NIH HHS · R01 CA087660 · United States
NIGMS NIH HHS · R01 GM090294-02 · United States
Databases
PDB
1ORI
External Links
PubMed source DOI: 10.1038/nature09472 PMC Full Text View journal details
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