Insights into the molecular recognition of the granuphilin C2A domain with PI(4,5)P2

Chanjuan Wan, Bo Wu, Zhenwei Song, Jiahai Zhang, Huiying Chu, Aoli Wang, Qingsong Liu, Yunyu Shi, Guohui Lic, Junfeng Wang

Chemistry and Physics of Lipids
2015 vol: 186 pp: 61-67 doi: 10.1016/j.chemphyslip.2015.01.003

Abstract

Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is a key player in regulating the process of excytosis, including insulin secretion. Granuphilin, a tandem C2 domain containing protein, mediates the docking of insulin granules onto plasma membrane. The C2A domain plays key roles in this process through interaction with PI(4,5)P2. In this study, we have investigated the molecular recognition mechanism of granuphilin-C2A domain to PI(4,5)P2 head group, and further to PI(4,5)P2-nanodisc by NMR, ITC, MST and SEC methods. Our results demonstrate that PI(4,5)P2 binds to the concave surface of granuphilin-C2A domain. The key residues involved in the binding were validated by mutation analysis.

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Topics: ITC, Membrane proteins, Nanodiscs, NMR, Monolith – MicroScale Thermophoresis, MST, Proteins, Publications