2009 Fiscal Year Final Research Report

Comprehensive analysis of membrane protein interactions by using in vivo photo-cross-linking technology.

Research Project

Project/Area Number	20770148
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Single-year Grants
Research Field	Molecular biology
Research Institution	The Institute of Physical and Chemical Research
Principal Investigator	HINO Nobumasa The Institute of Physical and Chemical Research, 拡張遺伝暗号システム研究チーム, 研究員 (90469916)
Project Period (FY)	2008 – 2009
Keywords	蛋白質間相互作用 / 膜タンパク質 / クロスリンク / 拡張遺伝暗号 / 非天然型アミノ酸
Research Abstract	To investigate the in vivo interaction between GPCRs and membrane proteins by a photo-cross-linking approach, a cross-linkable, non-natural amino acid was site-specifically incorporated at various positions of the extra-cellular domain of receptor-activity modified protein 1 (RAMP1) in mammalian cells. By exposing the cells to the light at 365 nm, the RAMP1 variants, each containing the non-natural amino acid at a specific site, were found to be efficiently cross-linked with the calcitonin receptor-like receptor, which is a GPCR known to interact with RAMP1. These variants were able to be cross-linked also with another GPCR, the calcitonin receptor. This observation revealed that RAMP1 binds to these different types of GPCRs directly, and the binding mode hardly changes, depending on the type of GPCR.