US 2022/0260583 A1 · CHEMOPROTEOMIC PROFILING OF PROTEIN ELECTROPHILIC AND OXIDATIVE POST-TRANSLATIONAL MODIFICATIONS

Abstract

Chemoproteomic methods for detecting and profiling electrophilic post-translational modifications (PTMs) and oxidative PTMs in proteins are described. The methods including contacting a proteomic mixture with a probe having hydrazine and an affinity handle to form a covalent linkage between the hydrazine moiety of the probe and the endogenous electrophilic PTM or the endogenous oxidative PTM. The resulting derivatized proteins are labelled with an tag via a click chemistry reaction. The labelled proteins can then be detected or profiled using techniques such as, for example, fluorescence imaging or mass spectrometry. Also described are protein conjugates having a covalent linkage formed by reaction of a hydrazine or oxyamine moiety of a probe with an electrophilic or oxidative PTM of a protein.

Claims19

Dependent claims

A method for detecting an endogenous electrophilic post-translational modification (PTM) or an endogenous oxidative PTM of at least one protein in a proteomic mixture in a living organism or living cell, the method comprising:

contacting the proteomic mixture in the living organism or living cell with a probe comprising a hydrazine moiety and an affinity handle to form a covalent linkage between the hydrazine moiety of the probe and the endogenous electrophilic PTM or the endogenous oxidative PTM of the at least one protein, thereby forming a mixture comprising at least one derivatized protein, wherein the endogenous electrophilic PTM or the endogenous oxidative PTM is installed onto the at least one protein by dedicated enzyme machinery or by high affinity binding of metabolite-derived small molecules;
labelling the at least one derivatized protein with a tag by conjugating the tag to the affinity handle to obtain at least one labelled protein;
and
analyzing the at least one labelled protein to thereby detect the endogenous electrophilic PTM or the endogenous oxidative PTM of the at least one protein.

1 more element

Dependencies are inferred from the claim text.

Chemoproteomic profiling of protein electrophilic and oxidative post-translational modifications

Life of this patent

Chemoproteomic profiling of protein electrophilic and oxidative post-translational modifications

Life of this patent

Abstract

Claims19