Method for the selective enrichment and labeling of phosphorproteins

摘要

The embodiments of the invention relate to a method for the introduction of a labeling structure such as a fluorescent molecules or a Raman tags to a compound. Imidazole functionalized resins or polymers are used to selectively immobilize phosphocompounds without protecting the carboxylic groups. Relying on the pKa difference between amines and hydrazides and carrying out the reaction in a slightly acidic buffer, all of the amines are protected by protonation while the hydrazides react with the phosphate imidazolide to form a phosphoramidate bond.

主权项

1. A method comprising:
reacting a phosphocompound having an intermediate compound attached thereto with a derivatization compound under a pH condition in which the derivatization compound has a lower acid ionization constant than the intermediate compound, wherein the derivatization compound has the formula R1R2—N—N—R3R4 and wherein at least one of R1-R4 is an acyl group, and wherein at least one of R1-R4 is a label, wherein the phosphocompound comprises phosphopeptide, phosphoprotein, phosphonucleotide or phosphooligonucleotide, and wherein the intermediate compound is an imidazole resin.