Phosphorylation-driven protein-protein interactions: a protein kinase sensing system.
A highly flexible protein kinase sensing system is described that furnishes severalfold changes in fluorescence in response to phosphorylation. A library of Src kinase peptide substrates was prepared that contained different environmentally sensitive fluorophores positioned at various sites on the active site directed sequence. Robust changes in fluorescent intensity were observed in the presence of a phosphotyrosine binding domain protein (Lck SH2 domain), which furnishes a hydrophobic environment for the fluorophore. This protein kinase sensing system has the advantages that the fluorescent indicator can be unobtrusively positioned on the peptide substrate, and that different environmentally sensitive fluorophores with distinct photophysical properties can be employed.
Wang
Qunzhao
Q
Department of Biochemistry, The Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
Lawrence
David S
DS
D000613:Aminobutyrates / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D002384:Catalysis; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000737:chemistry / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism; D010748:Phosphopeptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000032:analysis* / Q000378:metabolism; D018909:src Homology Domains
D000613:Aminobutyrates; D005456:Fluorescent Dyes; D009842:Oligopeptides; D010748:Phosphopeptides; C005959:2,4-diaminobutyric acid; D011494:Protein Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)