2011 Fiscal Year Final Research Report

Analysis of translation mechanism for selenoprotein synthesis

Research Project

Project/Area Number	22770093
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Single-year Grants
Research Field	Structural biochemistry
Research Institution	Hokkaido University
Principal Investigator	OSE Toyoyuki 北海道大学, 大学院・薬学研究院, 准教授 (80380525)
Project Period (FY)	2010 – 2011
Keywords	翻訳マシーナリ / RNA調製 / 蛋白質調製
Research Abstract	We successfully prepared protein and RNA molecules which are required for selecocystein incorporation on ribosome. These molecules are SBP2, L30, EFSec, SEICIS-RNA, tRNASec and subjected to biochemical and biophysical experiments. Crystal structure of L30 could be analyzed and published. Since we originally succeeded generating functional tRNASec, binding assay among tRNASec, EFSec, and SBP2 in the presence of specific amino acid are now in progress. We observed the significant binding affinity(K_d=229 nM) between the wild type and GDP using ITC, the way to prepare the molecule was thus confirmed. Furthermore, mutants generated for the purpose of crystallization were proved that they have proper secondary structures