title: Insights into the molecular recognition of the granuphilin C2A domain with PI(4,5)P2.
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.
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, PR China.
authors.forename: Chanjuan
authors.initials: C
authors.lastname: Wan
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, PR China. Electronic address: wubo@hmfl.ac.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Wu
authors.affiliation: Heifei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei 230027, PR China.
authors.forename: Zhenwei
authors.initials: Z
authors.lastname: Song
authors.affiliation: Heifei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei 230027, PR China.
authors.forename: Jiahai
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Laboratory of Molecular Modeling and Design, State key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Rd., Dalian 116023, PR China.
authors.forename: Huiying
authors.initials: H
authors.lastname: Chu
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, PR China.
authors.forename: Aoli
authors.initials: A
authors.lastname: Wang
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, PR China.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Heifei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei 230027, PR China.
authors.forename: Yunyu
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Laboratory of Molecular Modeling and Design, State key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Rd., Dalian 116023, PR China.
authors.forename: Guohui
authors.initials: G
authors.lastname: Li
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, PR China. Electronic address: junfeng@hmfl.ac.cn.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D006801:Humans; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D017434:Protein Structure, Tertiary; D033921:Vesicular Transport Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D018129:Phosphatidylinositol Phosphates; C512779:SYTL4 protein, human; D033921:Vesicular Transport Proteins; C060980:phosphatidylinositol 3,4-diphosphate
keywords: C2 domain; Granuphilin; NMR; Nanodisc; PI(4,5)P2; Recognition mechanism