RT Journal Article
SR Electronic
T1 Systematic identification of ALK substrates by integrated phosphoproteome and interactome analysis
JF Life Science Alliance
JO Life Sci. Alliance
FD Life Science Alliance LLC
SP e202101202
DO 10.26508/lsa.202101202
VO 5
IS 8
A1 Adachi, Jun
A1 Kakudo, Akemi
A1 Takada, Yoko
A1 Isoyama, Junko
A1 Ikemoto, Narumi
A1 Abe, Yuichi
A1 Narumi, Ryohei
A1 Muraoka, Satoshi
A1 Gunji, Daigo
A1 Hara, Yasuhiro
A1 Katayama, Ryohei
A1 Tomonaga, Takeshi
YR 2022
UL http://www.life-science-alliance.org/content/5/8/e202101202.abstract
AB The sensitivity of phosphorylation site identification by mass spectrometry has improved markedly. However, the lack of kinase–substrate relationship (KSR) data hinders the improvement of the range and accuracy of kinase activity prediction. In this study, we aimed to develop a method for acquiring systematic KSR data on anaplastic lymphoma kinase (ALK) using mass spectrometry and to apply this method to the prediction of kinase activity. Thirty-seven ALK substrate candidates, including 34 phosphorylation sites not annotated in the PhosphoSitePlus database, were identified by integrated analysis of the phosphoproteome and crosslinking interactome of HEK 293 cells with doxycycline-induced ALK overexpression. Furthermore, KSRs of ALK were validated by an in vitro kinase assay. Finally, using phosphoproteomic data from ALK mutant cell lines and patient-derived cells treated with ALK inhibitors, we found that the prediction of ALK activity was improved when the KSRs identified in this study were used instead of the public KSR dataset. Our approach is applicable to other kinases, and future identification of KSRs will facilitate more accurate estimations of kinase activity and elucidation of phosphorylation signals.