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Poster Presentations
Day 4, June 25(Wed.)
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-AM-47(2C-O3-1640)
Proteome-Wide Profiling of Protein Structural Dynamics by Phospho-Probing with Multiple Kinases
(1Kyoto Univ., 2NCVC, 3NIBIOHN)
oAsato Maeda1, Kosuke Ogata1, Naoyuki Sugiyama1,2, Yasushi Ishihama1,3
MS-based structural proteomics are increasingly being utilized to obtain a system-wide landscape of protein higher-order structural dynamics. Here we developed a novel phospho-probing approach utilizing multiple kinases to enhance the analytical coverage of protein structural changes. Protein kinases exhibited distinct substrate preferences, influencing phospho-probing efficiency in non-denatured versus structure-disrupted, trypsin-digested samples. Notably, 21.3 % of phosphorylation sites in trypsin-digested samples were uniquely targeted by a single kinase, compared to 13.0 % in non-denatured samples. These findings confirmed that many sites exposed upon structural disruption were selectively phosphorylated by specific kinases, and the use of multiple kinases improved the comprehensiveness of structural analysis with phospho-probing.