2013 Fiscal Year Final Research Report

Identification of cellular substrate for CaMKII using unnatural amino acid incorporation

Research Project

Project/Area Number	24650204
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Single-year Grants
Research Field	Neurochemistry/Neuropharmacology
Research Institution	Institute of Physical and Chemical Research
Principal Investigator	SANEYOSHI Takeo 独立行政法人理化学研究所, 脳科学総合研究センター, 研究員 (00556201)
Project Period (FY)	2012-04-01 – 2014-03-31
Keywords	シナプス / CaMKII / 酵素 / 非天然アミノ酸
Research Abstract	It is widely accepted that Calmodulin-kinase II (CaMKII) plays important roles in learning and memory. Although plasticity of synapse regulated by CaMKII, direct targets of CaMKII has poorly identified. To obtain cellular CaMKII substrates, we tried to apply unnatural amino acid incorporation method. Several phenylalanine residues in catalytic domain of CaMKII were made for subsequent incorporation of an unnatural amino acid pBpa. The pBpa were successfully incorporated on CaMKII in HEK293 cells. The covalent bonds were formed upon UV irradiation in cells expressing CaMKII mutants, especially residues, F16, F157, F171. This mutants are useful molecules to identify cellular substrate for CaMKII.