Combinatorial peptide microarrays for the rapid determination of kinase specificity.
We report a rapid method for profiling of kinases using a strategy that couples the merits of combinatorics (in rapid diversity generation) with the throughput attainable using microarrays (in parallel screening). Alanine-scanning, deletion and positional-scanning peptide libraries of a kinase substrate were synthesized and site-specifically arrayed onto glass slides. The phosphorylation pattern of target sequences detected using fluorescently-labeled antiphosphoamino acid antibodies revealed the substrate preference of the kinase through its activity profile.
Uttamchandani
Mahesh
M
Department of Biological Sciences, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
Chan
Elaine W S
EW
Chen
Grace Y J
GY
Yao
Shao Q
SQ
D000595:Amino Acid Sequence; D000906:Antibodies; D020650:Combinatorial Chemistry Techniques; D007118:Immunoassay; D015335:Molecular Probes; D019151:Peptide Library; D019606:Phosphoamino Acids / Q000276:immunology; D010766:Phosphorylation; D010770:Phosphotransferases / Q000737:chemistry / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D013379:Substrate Specificity
D000906:Antibodies; D015335:Molecular Probes; D019151:Peptide Library; D019606:Phosphoamino Acids; D010770:Phosphotransferases