The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles

Sowmya V. Yelamanchili, Clemens Reisinger, Anja Becher, Stefan Sikorra, Hans Bigalke, Thomas Binz, Gudrun Ahnert-Hilger

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Synaptophysin and synaptobrevin are abundant membrane proteins of neuronal small synaptic vesicles. In mature, differentiated neurons they form the synaptophysin/synaptobrevin (Syp/Syb) complex. Synaptobrevin also interacts with the plasma membrane-associated proteins syntaxin and SNAP25, thereby forming the SNARE complex necessary for exocytotic membrane fusion. The two complexes are mutually exclusive. Synaptobrevin is a C-terminally membrane-anchored protein with one transmembrane domain. While its interaction with its SNARE partners is mediated exclusively by its N-terminal cytosolic region it has been unclear so far how binding to synaptophysin is accomplished. Here, we show that synaptobrevin can be cleaved in its synaptophysin-bound form by tetanus toxin and botulinum neurotoxin B, or by botulinum neurotoxin D, leaving shorter or longer C-terminal peptide chains bound to synaptophysin, respectively. A recombinant, C-terminally His-tagged synaptobrevin fragment bound to nickel beads specifically bound synaptophysin, syntaxin and SNAP25 from vesicular detergent extracts. After cleavage by tetanus toxin or botulinum toxin D light chain, the remaining C-terminal fragment no longer interacted with syntaxin or SNAP 25. In contrast, synaptophysin was still able to bind to the residual C-terminal synaptobrevin cleavage product. In addition, the His-tagged C-terminal synaptobrevin peptide 68-116 was also able to bind synaptophysin in detergent extracts from adult brain membranes. These data suggest that synaptophysin interacts with the C-terminal transmembrane part of synaptobrevin, thereby allowing the N-terminal cytosolic chain to interact freely with the plasma membrane-associated SNARE proteins. Thus, by binding synaptobrevin, synaptophysin may positively modulate neurotransmission.

Original languageEnglish (US)
Pages (from-to)467-475
Number of pages9
JournalEuropean Journal of Cell Biology
Volume84
Issue number4
DOIs
StatePublished - Apr 22 2005

Fingerprint

R-SNARE Proteins
Synaptophysin
Synaptic Vesicles
Qa-SNARE Proteins
SNARE Proteins
Membrane Proteins
Tetanus Toxin
Detergents
Blood Proteins
Cell Membrane
Peptides
Membrane Fusion
Nickel
Synaptic Transmission

Keywords

  • Clostridial neurotoxins
  • SNARE complex
  • Synaptobrevin
  • Synaptophysin
  • Syp/Syb complex

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

Cite this

The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles. / Yelamanchili, Sowmya V.; Reisinger, Clemens; Becher, Anja; Sikorra, Stefan; Bigalke, Hans; Binz, Thomas; Ahnert-Hilger, Gudrun.

In: European Journal of Cell Biology, Vol. 84, No. 4, 22.04.2005, p. 467-475.

Research output: Contribution to journalArticle

Yelamanchili, Sowmya V. ; Reisinger, Clemens ; Becher, Anja ; Sikorra, Stefan ; Bigalke, Hans ; Binz, Thomas ; Ahnert-Hilger, Gudrun. / The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles. In: European Journal of Cell Biology. 2005 ; Vol. 84, No. 4. pp. 467-475.
@article{f3e9a6b41c8b4682ae1295f202ae94b4,
title = "The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles",
abstract = "Synaptophysin and synaptobrevin are abundant membrane proteins of neuronal small synaptic vesicles. In mature, differentiated neurons they form the synaptophysin/synaptobrevin (Syp/Syb) complex. Synaptobrevin also interacts with the plasma membrane-associated proteins syntaxin and SNAP25, thereby forming the SNARE complex necessary for exocytotic membrane fusion. The two complexes are mutually exclusive. Synaptobrevin is a C-terminally membrane-anchored protein with one transmembrane domain. While its interaction with its SNARE partners is mediated exclusively by its N-terminal cytosolic region it has been unclear so far how binding to synaptophysin is accomplished. Here, we show that synaptobrevin can be cleaved in its synaptophysin-bound form by tetanus toxin and botulinum neurotoxin B, or by botulinum neurotoxin D, leaving shorter or longer C-terminal peptide chains bound to synaptophysin, respectively. A recombinant, C-terminally His-tagged synaptobrevin fragment bound to nickel beads specifically bound synaptophysin, syntaxin and SNAP25 from vesicular detergent extracts. After cleavage by tetanus toxin or botulinum toxin D light chain, the remaining C-terminal fragment no longer interacted with syntaxin or SNAP 25. In contrast, synaptophysin was still able to bind to the residual C-terminal synaptobrevin cleavage product. In addition, the His-tagged C-terminal synaptobrevin peptide 68-116 was also able to bind synaptophysin in detergent extracts from adult brain membranes. These data suggest that synaptophysin interacts with the C-terminal transmembrane part of synaptobrevin, thereby allowing the N-terminal cytosolic chain to interact freely with the plasma membrane-associated SNARE proteins. Thus, by binding synaptobrevin, synaptophysin may positively modulate neurotransmission.",
keywords = "Clostridial neurotoxins, SNARE complex, Synaptobrevin, Synaptophysin, Syp/Syb complex",
author = "Yelamanchili, {Sowmya V.} and Clemens Reisinger and Anja Becher and Stefan Sikorra and Hans Bigalke and Thomas Binz and Gudrun Ahnert-Hilger",
year = "2005",
month = "4",
day = "22",
doi = "10.1016/j.ejcb.2004.11.007",
language = "English (US)",
volume = "84",
pages = "467--475",
journal = "European Journal of Cell Biology",
issn = "0171-9335",
publisher = "Urban und Fischer Verlag GmbH und Co. KG",
number = "4",

}

TY - JOUR

T1 - The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles

AU - Yelamanchili, Sowmya V.

AU - Reisinger, Clemens

AU - Becher, Anja

AU - Sikorra, Stefan

AU - Bigalke, Hans

AU - Binz, Thomas

AU - Ahnert-Hilger, Gudrun

PY - 2005/4/22

Y1 - 2005/4/22

N2 - Synaptophysin and synaptobrevin are abundant membrane proteins of neuronal small synaptic vesicles. In mature, differentiated neurons they form the synaptophysin/synaptobrevin (Syp/Syb) complex. Synaptobrevin also interacts with the plasma membrane-associated proteins syntaxin and SNAP25, thereby forming the SNARE complex necessary for exocytotic membrane fusion. The two complexes are mutually exclusive. Synaptobrevin is a C-terminally membrane-anchored protein with one transmembrane domain. While its interaction with its SNARE partners is mediated exclusively by its N-terminal cytosolic region it has been unclear so far how binding to synaptophysin is accomplished. Here, we show that synaptobrevin can be cleaved in its synaptophysin-bound form by tetanus toxin and botulinum neurotoxin B, or by botulinum neurotoxin D, leaving shorter or longer C-terminal peptide chains bound to synaptophysin, respectively. A recombinant, C-terminally His-tagged synaptobrevin fragment bound to nickel beads specifically bound synaptophysin, syntaxin and SNAP25 from vesicular detergent extracts. After cleavage by tetanus toxin or botulinum toxin D light chain, the remaining C-terminal fragment no longer interacted with syntaxin or SNAP 25. In contrast, synaptophysin was still able to bind to the residual C-terminal synaptobrevin cleavage product. In addition, the His-tagged C-terminal synaptobrevin peptide 68-116 was also able to bind synaptophysin in detergent extracts from adult brain membranes. These data suggest that synaptophysin interacts with the C-terminal transmembrane part of synaptobrevin, thereby allowing the N-terminal cytosolic chain to interact freely with the plasma membrane-associated SNARE proteins. Thus, by binding synaptobrevin, synaptophysin may positively modulate neurotransmission.

AB - Synaptophysin and synaptobrevin are abundant membrane proteins of neuronal small synaptic vesicles. In mature, differentiated neurons they form the synaptophysin/synaptobrevin (Syp/Syb) complex. Synaptobrevin also interacts with the plasma membrane-associated proteins syntaxin and SNAP25, thereby forming the SNARE complex necessary for exocytotic membrane fusion. The two complexes are mutually exclusive. Synaptobrevin is a C-terminally membrane-anchored protein with one transmembrane domain. While its interaction with its SNARE partners is mediated exclusively by its N-terminal cytosolic region it has been unclear so far how binding to synaptophysin is accomplished. Here, we show that synaptobrevin can be cleaved in its synaptophysin-bound form by tetanus toxin and botulinum neurotoxin B, or by botulinum neurotoxin D, leaving shorter or longer C-terminal peptide chains bound to synaptophysin, respectively. A recombinant, C-terminally His-tagged synaptobrevin fragment bound to nickel beads specifically bound synaptophysin, syntaxin and SNAP25 from vesicular detergent extracts. After cleavage by tetanus toxin or botulinum toxin D light chain, the remaining C-terminal fragment no longer interacted with syntaxin or SNAP 25. In contrast, synaptophysin was still able to bind to the residual C-terminal synaptobrevin cleavage product. In addition, the His-tagged C-terminal synaptobrevin peptide 68-116 was also able to bind synaptophysin in detergent extracts from adult brain membranes. These data suggest that synaptophysin interacts with the C-terminal transmembrane part of synaptobrevin, thereby allowing the N-terminal cytosolic chain to interact freely with the plasma membrane-associated SNARE proteins. Thus, by binding synaptobrevin, synaptophysin may positively modulate neurotransmission.

KW - Clostridial neurotoxins

KW - SNARE complex

KW - Synaptobrevin

KW - Synaptophysin

KW - Syp/Syb complex

UR - http://www.scopus.com/inward/record.url?scp=16244419076&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=16244419076&partnerID=8YFLogxK

U2 - 10.1016/j.ejcb.2004.11.007

DO - 10.1016/j.ejcb.2004.11.007

M3 - Article

C2 - 15900706

AN - SCOPUS:16244419076

VL - 84

SP - 467

EP - 475

JO - European Journal of Cell Biology

JF - European Journal of Cell Biology

SN - 0171-9335

IS - 4

ER -