Crystallography of membrane protein complex by S-SAD method

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K14468
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: High Energy Accelerator Research Organization
• Principal Investigator: KATO Ryuichi 大学共同利用機関法人高エネルギー加速器研究機構, 物質構造科学研究所, 准教授 (50240833)
• Co-Investigator(Renkei-kenkyūsha): MAKIYO Hisayoshi 高エネルギー加速器研究機構, 物質構造科学研究所, 研究員 (00467474) ; MATSUGAKI Naohiro 高エネルギー加速器研究機構, 物質構造科学研究所, 准教授 (50342598) ; YAMADA Yusuke 高エネルギー加速器研究機構, 物質構造科学研究所, 助教 (20391708) ; MANYA Hiroshi 東京都健康長寿医療センター研究所, 研究副部長 (20321870)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 筋ジストロフィー / ジストログリカン / 膜タンパク質 / 結晶構造解析 / POMT1 / POMT2

Outline of Final Research Achievements

For structural study of the membrane protein complex POMT1-POMT2, which is involved in congenital muscular dystrophy, expression experiments in mammalian cultured cells were performed. Unfortunately, the expression level was very low, which is insufficient to start crystallization experiments. We also examined the method of crystallizing proteins expressed in insect body, but the proteins did not express.
In parallel, we studied development of a method of performing X-ray crystal structure analysis of membrane protein POMT1-POMT2 by a phase determination method utilizing the anomalous dispersion effect of sulfur atoms (S-SAD method). By using standard proteins with synchrotron beamline at KEK, S-SAD method was improved, and sufficient results were obtained.

Free Research Field

構造生物学