RRC ID 48556
Author Fukuda M, Kojima T, Aruga J, Niinobe M, Mikoshiba K.
Title Functional diversity of C2 domains of synaptotagmin family. Mutational analysis of inositol high polyphosphate binding domain.
Journal J Biol Chem
Abstract Synaptotagmins I and II are inositol high polyphosphate series (inositol 1,3,4,5-tetrakisphosphate (IP4), inositol 1,3,4,5,6-pentakisphosphate, and inositol 1,2,3,4,5,6-hexakisphosphate) binding proteins, which are thought to be essential for Ca(2+)-regulated exocytosis of neurosecretory vesicles. In this study, we analyzed the inositol high polyphosphate series binding site in the C2B domain by site-directed mutagenesis and compared the IP4 binding properties of the C2B domains of multiple synaptotagmins (II-IV). The IP4 binding domain of synaptotagmin II is characterized by a cluster of highly conserved, positively charged amino acids (321 GKRLKKKKTTVKKK 324). Among these, three lysine residues, at positions 327, 328, and 332 in the middle of the C2B domain, which is not conserved in the C2A domain, were found to be essential for IP4 binding in synaptotagmin II. When these lysine residues were altered to glutamine, the IP4 binding ability was completely abolished. The primary structures of the IP4 binding sites are highly conserved among synaptotagmins I through IV. However, synaptotagmin III did not show significant binding ability, which may be due to steric hindrance by the C-terminal flanking region. These functional diversities of C2B domains suggest that not all synaptotagmins function as inositol high polyphosphate sensors at the synaptic vesicle.
Volume 270(44)
Pages 26523-7
Published 1995-11-3
DOI 10.1074/jbc.270.44.26523
PMID 7592870
MeSH Amino Acid Sequence Animals Antibodies Base Sequence Binding Sites Calcium-Binding Proteins* Cloning, Molecular Conserved Sequence DNA Primers Genetic Variation* Inositol Phosphates / metabolism* Kinetics Membrane Glycoproteins / biosynthesis Membrane Glycoproteins / chemistry* Membrane Glycoproteins / metabolism* Mice Molecular Sequence Data Multigene Family Mutagenesis, Site-Directed Nerve Tissue Proteins / biosynthesis Nerve Tissue Proteins / chemistry* Nerve Tissue Proteins / metabolism* Rabbits / immunology Recombinant Fusion Proteins / metabolism Recombinant Proteins / biosynthesis Recombinant Proteins / chemistry Recombinant Proteins / metabolism Sequence Homology, Amino Acid Synaptotagmin II Synaptotagmins
IF 4.106
Times Cited 164
DNA material pEF-T7-Syt III (Syt3) (RDB14683) pEF-T7-Syt IV (Syt4) (RDB14684).