Methods to assay kinase activity

摘要

Methods to assay kinase activity are provided herein. The methods employ elemental analysis, including inductively charged plasma mass spectrometry (ICP-MS). The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by kinase enzymes involved in post-translational modifications.

主权项

1. A method to assay kinase activity, the method comprising:
a) incubating ATP, at least one kinase, and a free non-phosphorylated substrate labeled with an element tag, with a support having attached thereto a metal ion coordination complex under conditions to enable the kinase to phosphorylate the substrate; b) separating free non-phosphorylated substrate from phosphorylated substrate labeled with an element tag bound to the support; c) eluting the element tag associated with the resultant phosphorylated substrate into a solution; and d) performing solution elemental analysis of said solution by using inductively coupled plasma mass spectrometry to detect the element tag associated with the phosphorylated substrate.