title: A chemoproteomic platform to quantitatively map targets of lipid-derived electrophiles.
abstract: Cells produce electrophilic products with the potential to modify and affect the function of proteins. Chemoproteomic methods have provided a means to qualitatively inventory proteins targeted by endogenous electrophiles; however, ascertaining the potency and specificity of these reactions to identify the sites in the proteome that are most sensitive to electrophilic modification requires more quantitative methods. Here we describe a competitive activity-based profiling method for quantifying the reactivity of electrophilic compounds against >1,000 cysteines in parallel in the human proteome. Using this approach, we identified a select set of proteins that constitute 'hot spots' for modification by various lipid-derived electrophiles, including the oxidative stress product 4-hydroxy-2-nonenal (HNE). We show that one of these proteins, ZAK kinase, is labeled by HNE on a conserved, active site-proximal cysteine and that the resulting enzyme inhibition creates a negative feedback mechanism that can suppress the activation of JNK pathways normally induced by oxidative stress.
authors.affiliation: 1] The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA. [2] Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Chu
authors.initials: C
authors.lastname: Wang
authors.affiliation: 1] The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA. [2] Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Eranthie
authors.initials: E
authors.lastname: Weerapana
authors.affiliation: 1] The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA. [2] Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Megan M
authors.initials: MM
authors.lastname: Blewett
authors.affiliation: 1] The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA. [2] Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
mesh_terms: D000595:Amino Acid Sequence; D001943:Breast Neoplasms / Q000378:metabolism; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D003545:Cysteine / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004563:Electrochemistry / Q000379:methods; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D008055:Lipids / Q000737:chemistry; D020930:MAP Kinase Kinase Kinases; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D015230:Prostaglandin D2 / Q000031:analogs & derivatives / Q000737:chemistry; D011494:Protein Kinases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D017386:Sequence Homology, Amino Acid
chemical_list: C477819:15-deoxyprostaglandin J2; D008055:Lipids; D020543:Proteome; D011494:Protein Kinases; D020930:MAP Kinase Kinase Kinases; C413047:MAP3K20 protein, human; D003545:Cysteine; D015230:Prostaglandin D2