Phosphorylation-protein complementation assay and related systems and methods

Abstract

Provided herein include a phosphorylation-protein complementation reporter system, an abundance-protein complementation reporter systems, and assays and methods of using the same to detect phosphorylation and for high-throughput kinase variant screening. In some embodiments, the phosphorylation-protein complementation reporter system comprises a first fusion protein comprising a kinase substrate linked to (i) a kinase domain and (ii) a first segment of a reporter molecule; and a second fusion protein comprising a phosphor-amino-acid binding domain (PAABD) linked to a second segment of the reporter molecule, wherein the first segment and the second segment of the reporter molecule are capable of complementing one another to form an active reporter molecule via the interaction between the PAABD and the kinase substrate upon phosphorylation by the kinase domain.

Dependencies are inferred from the claim text.