Structural analysis of cell adhesion molecules that determine cell coordinate in tissue

2016 Fiscal Year Final Research Report

• Project/Area Number: 26440028
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Osaka University
• Principal Investigator: Suzuki Mamoru 大阪大学, たんぱく質研究所, 准教授 (40280507)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ネクチン / 構造生物学 / 結晶構造解析

Outline of Final Research Achievements

We established recombinant expression systems for extracellular domains of mouse Nectin-1 and mouse Nectin-3 in insect cell Sf-9. First we tried to purify Nectin-1 / Nectin-3 heterodimer fraction from the mixture of cultural supernatants. However, the result of surface plasmon resonance analysis indicated the fast dissociation rate constant of heterophilic interaction between Nectin-1 and Nectin-3, suggested difficulty of stable hetero-complex purification. Thus we next established individual purification systems for Nectin-1 and Nectin-3, respectively. The purified Nectins were mixed and used for crystallization experiments. As a result, structures of Nectin-1 homodimer and Nectin-3 homodimer were obtained from mixture of purified Nectin-1 and Nectin-3 in 2.24 A and 2.58 A resolutions, respectively (PDB ID: 5B21and 5B22). Additionally, chemical crosslink experiment using BS3 crosslinker revealed that the minimal unit of Nectin-1 / Nectin-3 heterophilic complex is heterodimer.

Free Research Field

構造生物科学