Article: article from journal or magazin.
Direct interaction of the Rab3 effector RIM with Ca2+ channels, SNAP-25, and synaptotagmin.
Journal of Biological Chemistry
To define the role of the Rab3-interacting molecule RIM in exocytosis we searched for additional binding partners of the protein. We found that the two C(2) domains of RIM display properties analogous to those of the C(2)B domain of synaptotagmin-I. Thus, RIM-C(2)A and RIM-C(2)B bind in a Ca(2+)-independent manner to alpha1B, the pore-forming subunit of N-type Ca(2+) channels (EC(50) = approximately 20 nm). They also weakly interact with the alpha1C but not the alpha1D subunit of L-type Ca(2+) channels. In addition, the C(2) domains of RIM associate with SNAP-25 and synaptotagmin-I. The binding affinities for these two proteins are 203 and 24 nm, respectively, for RIM-C(2)A and 224 and 16 nm for RIM-C(2)B. The interactions of the C(2) domains of RIM with SNAP-25 and synaptotagmin-I are modulated by Ca(2+). Thus, in the presence of Ca(2+) (EC(50) = approximately 75 microm) the interaction with synaptotagmin-I is increased, whereas SNAP-25 binding is reduced. Synaptotagmin-I binding is abolished by mutations in two positively charged amino acids in the C(2) domains of RIM and by the addition of inositol polyphosphates. We propose that the Rab3 effector RIM is a scaffold protein that participates through its multiple binding partners in the docking and fusion of secretory vesicles at the release sites.
Amino Acid Sequence, Animals, Antigens, Surface/chemistry, Antigens, Surface/metabolism, Binding Sites, Brain/metabolism, Calcium/metabolism, Calcium/pharmacology, Calcium Channels/chemistry, Calcium Channels/metabolism, Calcium-Binding Proteins, Cloning, Molecular, GTP-Binding Proteins, Humans, Kinetics, Membrane Glycoproteins/chemistry, Membrane Glycoproteins/metabolism, Membrane Proteins/chemistry, Membrane Proteins/metabolism, Molecular Sequence Data, Mutagenesis, Site-Directed, Nerve Tissue Proteins/chemistry, Nerve Tissue Proteins/genetics, Protein Isoforms/chemistry, Protein Isoforms/genetics, Protein Subunits, RNA, Messenger/genetics, Rats, Recombinant Proteins/chemistry, Recombinant Proteins/metabolism, Sequence Alignment, Sequence Homology, Amino Acid, Synaptosomal-Associated Protein 25, Synaptotagmin I, Synaptotagmins, Syntaxin 1, Zinc Fingers, rab3 GTP-Binding Proteins/metabolism
Web of science
Last modification date