Program

Poster Presentations

Day 2, May 16（Wed.）　　Poster

2P-50（3B-O1-M-0915）　　PDF

Determination of Sequence Preference of Kinase Substrates Based on Large-Scale Identification of in vitro Substrates

(Kyoto Univ.)
^oJunna Nakazono, Dai Sakamoto, Hsin-yi Chang, Naoyuki Sugiyama, Yasushi Ishihama

Protein kinase is one of the key components in phosphorylation-based signal transduction, and aberrant kinase activity often results in severe diseases including cancer. Therefore, quantifying kinase activities is valuable for diagnostic and therapeutic purposes. While recent advances in phosphoproteomics with LC/MS enable us to identify thousands of phosphosites, it is still challenging to obtain kinome activity profiles because the information of kinase-substrate relationship is mostly missing. Previously, we designed substrate peptides with high specificity and high sensitivity to each 187 serine/threonine kinase, and the experimental confirmation by in vitro kinase reaction for each kinase indicated that more substrate information was needed to increase both specificity and sensitivity in some cases. In this study, we performed deeper phosphoproteomics analysis of the lysate proteins reacted with recombinant kinase in vitro to identify as many substrates as possible for determining kinase-specific sequence preference of substrates.