Cysteine reactive probes and uses thereof
AssigneeThe Scripps Research Institute
InventorsBenjamin F. CRAVATT, Keriann M. BACKUS, Bruno E. CORREIA, Megan M. BLEWETT, John R. TEIJARO
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Abstract
Disclosed herein are methods, compositions, probes, polypeptides, assays, and kits for identifying a cysteine containing protein as a binding target for a small molecule fragment. Also disclosed herein are methods, compositions, and probes for mapping a biologically active cysteine site on a protein and screening a small molecule fragment for interaction with a cysteine containing protein.
Claims30
A modified cysteine containing protein comprising a small molecule fragment having a covalent bond to a cysteine residue of a cysteine containing protein, wherein the small molecule fragment has a molecular weight of about 150 Dalton or higher.
A method of identifying a cysteine containing protein as a binding target for a small molecule fragment, comprising:
- a) obtaining a set of cysteine-reactive probe-protein complexes from a sample comprising a first cell solution treated with a small molecule fragment and a cysteine reactive probe wherein the cysteine-reactive probe comprises a reactive moiety capable of forming a covalent bond with a cysteine residue located on the cysteine containing protein;
- b) analyzing the set of cysteine-reactive probe-protein complexes by a proteomic analysis means;
- and
- c) based on step b), identifying a cysteine containing protein as the binding target for the small molecule fragment.
A method of screening a small molecule fragment for interaction with a cysteine containing protein, comprising:
- a) harvesting a set of cysteine-reactive probe-protein complexes from a sample comprising a first cell solution treated with a small molecule fragment and a cysteine reactive probe wherein the cysteine-reactive probe comprises a reactive moiety capable of forming a covalent bond with a cysteine residue located on the cysteine containing protein;
- b) analyzing the set of cysteine-reactive probe-protein complexes by a proteomic analysis means;
- and
- c) based on step b), identifying the small molecule fragment as interacting with the cysteine containing protein.
Dependencies are inferred from the claim text.