Vesicle-associated membrane protein-2/Synaptobrevin binding to Synaptotagmin I promotes O-glycosylation of Synaptotagmin I

被引:42
作者
Fukuda, M [1 ]
机构
[1] RIKEN, Inst Phys & Chem Res, Fukuda Initiat Res Unit, Wako, Saitama 3510198, Japan
关键词
D O I
10.1074/jbc.M204056200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Synaptotagmin I (Syt I), an evolutionarily conserved integral membrane protein of synaptic vesicles, is now known to regulate Ca2+-dependent neurotransmitter release. Syt I protein should undergo several post-translational modifications before maturation and subsequent functioning on synaptic vesicles (e.g. N-glycosylation and fatty acylation in vertebrate Syt I), because the apparent molecular weight of Syt I on synaptic vesicles (mature form, 65,000) was much higher than the calculated molecular weight (47,400) predicted from the cDNA sequences both in vertebrates and invertebrates. Common post-translational modification(s) of Syt I conserved across phylogeny, however, have never been elucidated. In the present study, I discovered that dithreonine residues (Thr-15 and Thr-16) at the intravesicular domain of mouse Syt I are post-translationally modified by a complex form of O-linked sugar (i.e. the addition of sialic acids) in PC12 cells and that the O-glycosylation of Syt I in COS-7 cells depends on the coexpression of vesicle-associated membrane protein-2 (VAMP-2)/synaptobrevin. I also showed that a transmembrane domain of Syt I directly interacts with isolated VAMP-2, but not VAMP-2, in the heterotrimeric SNARE (SNAP receptor) complex (vesicle SNARE, VAMP-2, and two target SNARES, syntaxin IA and SNAP-25). Since di-Thr or di-Ser residues are often found at the intravesicular domain of invertebrate Syt I, and VAMP-dependent O-glycosylation was also observed in squid Syt expressed in COS-7 cells, I propose that VAMP-dependent O-glycosylation of Syt I is a common modification during evolution and may have important role(s) in synaptic vesicle trafficking.
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页码:30351 / 30358
页数:8
相关论文
共 88 条
[1]   SYNAPTOBREVIN - AN INTEGRAL MEMBRANE-PROTEIN OF 18000 DALTONS PRESENT IN SMALL SYNAPTIC VESICLES OF RAT-BRAIN [J].
BAUMERT, M ;
MAYCOX, PR ;
NAVONE, F ;
DECAMILLI, P ;
JAHN, R .
EMBO JOURNAL, 1989, 8 (02) :379-384
[2]   SYNTAXIN - A SYNAPTIC PROTEIN IMPLICATED IN DOCKING OF SYNAPTIC VESICLES AT PRESYNAPTIC ACTIVE ZONES [J].
BENNETT, MK ;
CALAKOS, N ;
SCHELLER, RH .
SCIENCE, 1992, 257 (5067) :255-259
[3]   Synaptotagamin I and IV define distinct populations of neuronal transport vesicles [J].
Berton, F ;
Cornet, V ;
Iborra, C ;
Garrido, J ;
Dargent, B ;
Fukuda, M ;
Seagar, M ;
Marquèze, B .
EUROPEAN JOURNAL OF NEUROSCIENCE, 2000, 12 (04) :1294-1302
[4]   INHIBITION OF NEUROTRANSMITTER RELEASE BY C2-DOMAIN PEPTIDES IMPLICATES SYNAPTOTAGMIN IN EXOCYTOSIS [J].
BOMMERT, K ;
CHARLTON, MP ;
DEBELLO, WM ;
CHIN, GJ ;
BETZ, H ;
AUGUSTINE, GJ .
NATURE, 1993, 363 (6425) :163-165
[5]   The subcellular localizations of atypical synaptotagmins III and VI -: Synaptotagmin III is enriched in synapses and synaptic plasma membranes but not in synaptic vesicles [J].
Butz, S ;
Fernandez-Chacon, R ;
Schmitz, F ;
Jahn, R ;
Südhof, TC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (26) :18290-18296
[6]   Fatty acylation of synaptotagmin in PC12 cells and synaptosomes [J].
Chapman, ER ;
Blasi, J ;
An, S ;
Brose, N ;
Johnston, PA ;
Sudhof, TC ;
Jahn, R .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1996, 225 (01) :326-332
[7]   Alternative splicing of synaptotagmins involving transmembrane exon skipping [J].
Craxton, M ;
Goedert, M .
FEBS LETTERS, 1999, 460 (03) :417-422
[8]   Genomic analysis of synaptotagmin genes [J].
Craxton, M .
GENOMICS, 2001, 77 (1-2) :43-49
[9]   Kinetics of synaptotagmin responses to Ca2+ and assembly with the core SNARE complex onto membranes [J].
Davis, AF ;
Bai, JH ;
Fasshauer, D ;
Wolowick, MJ ;
Lewis, JL ;
Chapman, ER .
NEURON, 1999, 24 (02) :363-376
[10]   The C2B domain of synaptotagmin is a Ca2+-sensing module essential for exocytosis [J].
Desai, RC ;
Vyas, B ;
Earles, CA ;
Littleton, JT ;
Kowalchyck, JA ;
Martin, TFJ ;
Chapman, ER .
JOURNAL OF CELL BIOLOGY, 2000, 150 (05) :1125-1135