Home LiteratureArticle Details
PMID: 12939392 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Examining synaptotagmin 1 function in dense core vesicle exocytosis under direct control of Ca2+.

The Journal of general physiology ·Vol. 122 ·No. 3 ·2003-09-00 ·Pages 265-76

Sørensen JB, Fernández-Chacón R, Südhof TC, Neher E

Abstract

We tested the long-standing hypothesis that synaptotagmin 1 is the Ca2+ sensor for fast neurosecretion by analyzing the intracellular Ca2+ dependence of large dense-core vesicle exocytosis in a mouse strain carrying a mutated synaptotagmin C2A domain. The mutation (R233Q) causes a twofold increase in the KD of Ca2+-dependent phospholipid binding to the double C2A-C2B domain of synaptotagmin. Using photolysis of caged calcium and capacitance measurements we found that secretion from mutant cells had lower secretory rates, longer secretory delays, and a higher intracellular Ca2+-threshold for secretion due to a twofold increase in the apparent KD of the Ca2+ sensor for fast exocytosis. Single amperometric fusion events were unchanged. We conclude that Ca2+-dependent phospholipid binding to synaptotagmin 1 mirrors the intracellular Ca2+ dependence of exocytosis.

MeSH Terms
Animals Arginine/genetics Calcium/physiology Calcium-Binding Proteins Chromaffin Cells/metabolism Differential Threshold Electric Capacitance Exocytosis/physiology Glutamine/genetics Intracellular Membranes/metabolism Membrane Glycoproteins/genetics,physiology Mice Mice, Mutant Strains Nerve Tissue Proteins/genetics,physiology Phospholipids/metabolism Point Mutation/genetics Secretory Vesicles/physiology Synaptotagmin I Synaptotagmins Time Factors
Chemicals
Calcium-Binding Proteins Membrane Glycoproteins Nerve Tissue Proteins Phospholipids Synaptotagmin I Syt1 protein, mouse Glutamine Synaptotagmins Arginine Calcium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Sørensen Jakob B
Max-Planck-Institut für Biophysikalische Chemie, Abteilung Membranbiophysik, Am Fassberg 11, D-37077 Göttingen, Germany. jsoeren@gwdg.de
Fernández-Chacón Rafael
Südhof Thomas C
Neher Erwin
References (51)
51 references, click to expand
  1. Interaction of the synprint site of N-type Ca2+ channels with the C2B domain of synaptotagmin I.
    Proc Natl Acad Sci U S A. 1997 May 13;94(10):5405-10 PMID: 9144250
  2. Synaptotagmin V is targeted to dense-core vesicles that undergo calcium-dependent exocytosis in PC12 cells.
    J Biol Chem. 2002 Jul 5;277(27):24499-505 PMID: 12006594
  3. Synaptotagmin I: a major Ca2+ sensor for transmitter release at a central synapse.
    Cell. 1994 Nov 18;79(4):717-27 PMID: 7954835
  4. Three-dimensional structure of the synaptotagmin 1 C2B-domain: synaptotagmin 1 as a phospholipid binding machine.
    Neuron. 2001 Dec 20;32(6):1057-69 PMID: 11754837
  5. Syntaxin: a synaptic protein implicated in docking of synaptic vesicles at presynaptic active zones.
    Science. 1992 Jul 10;257(5067):255-9 PMID: 1321498
  6. Synaptic function is impaired but not eliminated in C. elegans mutants lacking synaptotagmin.
    Cell. 1993 Jul 2;73(7):1291-305 PMID: 8391930
  7. Synaptotagmin I functions as a calcium regulator of release probability.
    Nature. 2001 Mar 1;410(6824):41-9 PMID: 11242035
  8. Ca2+ regulates the interaction between synaptotagmin and syntaxin 1.
    J Biol Chem. 1995 Oct 6;270(40):23667-71 PMID: 7559535
  9. Intracellular calcium dependence of transmitter release rates at a fast central synapse.
    Nature. 2000 Aug 24;406(6798):889-93 PMID: 10972290
  10. Phospholipid binding by a synaptic vesicle protein homologous to the regulatory region of protein kinase C.
    Nature. 1990 May 17;345(6272):260-3 PMID: 2333096
  11. Synaptotagmin: a calcium sensor on the synaptic vesicle surface.
    Science. 1992 May 15;256(5059):1021-5 PMID: 1589771
  12. Synaptotagmin VII as a plasma membrane Ca(2+) sensor in exocytosis.
    Neuron. 2001 May;30(2):459-73 PMID: 11395007
  13. Dissection of three Ca2+-dependent steps leading to secretion in chromaffin cells from mouse adrenal slices.
    Neuron. 2000 Nov;28(2):537-45 PMID: 11144362
  14. Calcium dependence of exocytosis and endocytosis at the cochlear inner hair cell afferent synapse.
    Neuron. 2001 Mar;29(3):681-90 PMID: 11301027
  15. A novel function for the second C2 domain of synaptotagmin. Ca2+-triggered dimerization.
    J Biol Chem. 1996 Mar 8;271(10):5844-9 PMID: 8621455
  16. A single C2 domain from synaptotagmin I is sufficient for high affinity Ca2+/phospholipid binding.
    J Biol Chem. 1993 Dec 15;268(35):26386-90 PMID: 8253763
  17. The SNARE protein SNAP-25 is linked to fast calcium triggering of exocytosis.
    Proc Natl Acad Sci U S A. 2002 Feb 5;99(3):1627-32 PMID: 11830673
  18. Intracellular calcium dependence of large dense-core vesicle exocytosis in the absence of synaptotagmin I.
    Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11680-5 PMID: 11562488
  19. Synaptotagmin I functions as a calcium sensor to synchronize neurotransmitter release.
    Neuron. 2002 Dec 5;36(5):897-908 PMID: 12467593
  20. Synaptotagmin I is a high affinity receptor for clathrin AP-2: implications for membrane recycling.
    Cell. 1994 Sep 9;78(5):751-60 PMID: 8087843
  21. Nonuniform probability of glutamate release at a hippocampal synapse.
    Science. 1993 Oct 29;262(5134):754-7 PMID: 7901909
  22. The C terminus of SNAP25 is essential for Ca(2+)-dependent binding of synaptotagmin to SNARE complexes.
    J Biol Chem. 2000 Mar 3;275(9):6328-36 PMID: 10692432
  23. The reduced release probability of releasable vesicles during recovery from short-term synaptic depression.
    Neuron. 1999 Aug;23(4):821-32 PMID: 10482247
  24. Differential control of the releasable vesicle pools by SNAP-25 splice variants and SNAP-23.
    Cell. 2003 Jul 11;114(1):75-86 PMID: 12859899
  25. Synaptic depression and the kinetics of exocytosis in retinal bipolar cells.
    J Neurosci. 2000 Jan 15;20(2):568-78 PMID: 10632586
  26. Mechanisms underlying phasic and sustained secretion in chromaffin cells from mouse adrenal slices.
    Neuron. 1999 Jul;23(3):607-15 PMID: 10433271
  27. Functional diversity of C2 domains of synaptotagmin family. Mutational analysis of inositol high polyphosphate binding domain.
    J Biol Chem. 1995 Nov 3;270(44):26523-7 PMID: 7592870
  28. Ca2+-dependent synaptotagmin binding to SNAP-25 is essential for Ca2+-triggered exocytosis.
    Neuron. 2002 May 16;34(4):599-611 PMID: 12062043
  29. Ca2+-dependent and -independent interactions of the isoforms of the alpha1A subunit of brain Ca2+ channels with presynaptic SNARE proteins.
    Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14782-6 PMID: 9405690
  30. Co-operative action a calcium ions in transmitter release at the neuromuscular junction.
    J Physiol. 1967 Nov;193(2):419-32 PMID: 6065887
  31. Ca(2+)-dependent and -independent activities of neural and non-neural synaptotagmins.
    Nature. 1995 Jun 15;375(6532):594-9 PMID: 7791877
  32. Synaptotagmin III is a novel isoform of rat synaptotagmin expressed in endocrine and neuronal cells.
    J Biol Chem. 1994 Apr 22;269(16):11675-8 PMID: 8163462
  33. Calcium-dependent switching of the specificity of phosphoinositide binding to synaptotagmin.
    Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13327-32 PMID: 8917590
  34. Distinct Ca(2+)-dependent properties of the first and second C2-domains of synaptotagmin I.
    J Biol Chem. 1996 Jan 19;271(3):1262-5 PMID: 8576108
  35. Intrinsic voltage dependence and Ca2+ regulation of mslo large conductance Ca-activated K+ channels.
    J Gen Physiol. 1997 May;109(5):647-73 PMID: 9154910
  36. Synaptotagmin modulation of fusion pore kinetics in regulated exocytosis of dense-core vesicles.
    Science. 2001 Nov 2;294(5544):1111-5 PMID: 11691996
  37. Calcium dependence of the rate of exocytosis in a synaptic terminal.
    Nature. 1994 Oct 6;371(6497):513-5 PMID: 7935764
  38. Mutational analysis of Drosophila synaptotagmin demonstrates its essential role in Ca(2+)-activated neurotransmitter release.
    Cell. 1993 Sep 24;74(6):1125-34 PMID: 8104705
  39. Synaptotagmins form a hierarchy of exocytotic Ca(2+) sensors with distinct Ca(2+) affinities.
    EMBO J. 2002 Feb 1;21(3):270-80 PMID: 11823420
  40. Structure/function analysis of Ca2+ binding to the C2A domain of synaptotagmin 1.
    J Neurosci. 2002 Oct 1;22(19):8438-46 PMID: 12351718
  41. The C(2)B Ca(2+)-binding motif of synaptotagmin is required for synaptic transmission in vivo.
    Nature. 2002 Jul 18;418(6895):340-4 PMID: 12110842
  42. Quantitative and statistical analysis of the shape of amperometric spikes recorded from two populations of cells.
    J Neurochem. 2000 Mar;74(3):1086-97 PMID: 10693940
  43. A simple method for insulating carbon-fiber microelectrodes using anodic electrophoretic deposition of paint.
    Anal Chem. 1996 Sep 1;68(17):3054-8 PMID: 21619374
  44. Synaptotagmin IX regulates Ca2+-dependent secretion in PC12 cells.
    J Biol Chem. 2002 Feb 15;277(7):4601-4 PMID: 11751925
  45. How does calcium trigger neurotransmitter release?
    Curr Opin Neurobiol. 2001 Jun;11(3):320-6 PMID: 11399430
  46. Quantitative relationship between transmitter release and calcium current at the calyx of held synapse.
    J Neurosci. 2001 Jan 15;21(2):462-76 PMID: 11160426
  47. Synaptotagmin regulation of coated pit assembly.
    J Biol Chem. 2000 Oct 6;275(40):30916-24 PMID: 10906143
  48. Sr2+ binding to the Ca2+ binding site of the synaptotagmin 1 C2B domain triggers fast exocytosis without stimulating SNARE interactions.
    Neuron. 2003 Jan 9;37(1):99-108 PMID: 12526776
  49. Synaptotagmins I and IV promote transmitter release independently of Ca(2+) binding in the C(2)A domain.
    Nature. 2002 Jul 18;418(6895):336-40 PMID: 12110845
  50. Calcium-dependent interaction of the cytoplasmic region of synaptotagmin with membranes. Autonomous function of a single C2-homologous domain.
    J Biol Chem. 1994 Feb 25;269(8):5735-41 PMID: 8119912
  51. Calcium sensitivity of glutamate release in a calyx-type terminal.
    Science. 2000 Aug 11;289(5481):953-7 PMID: 10937999
Article Info
Journal
The Journal of general physiology
Abbr.
J Gen Physiol
ISSN
0022-1295
DOI
10.1085/jgp.200308855
Email
jsoeren@gwdg.de
Published
2003-09-00
Pages
265-76
Language
English
Region
United States
NLM ID
2985110R
PMCID
PMC2234490
Subset
IM
Corrections
CommentIn
PMID: 12939390
External Links
PubMed source DOI: 10.1085/jgp.200308855 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2