Vesicle-associated proteins and transmitter release from sympathetic ganglionic boutons.
A method is reported for introducing peptides derived from SNARE proteins that control exocytosis of vesicles at boutons formed by sympathetic ganglion cells in tissue culture. These peptides were coupled to the DNA binding domain of the Drosophila transcription factor antennapedia, called penetratin. This facilitated the passage of peptides across the bouton membrane. FM1-43 was used to monitor the exocytosis of transmitter from depolarized boutons after their exposure to the penetratin-peptide sequences IETRHNEIIKLETSIRELHD of syntaxin and KGFLSSLFGGSSK of alpha-SNAP, both of which blocked secretion, whereas the peptide sequences SELDDRA-DALQAGASQFETSAAKLKRK of synaptobrevin did not. This report introduces a readily applicable method for determining the effect of different peptide sequences of vesicle-associated proteins on secretion at vertebrate boutons and presents an account of the effects of a selection of such peptides on exocytosis.
Blair
D H
DH
Neurobiology Laboratory, University of Sydney, NSW, Australia.
Robson
S
S
King
G
G
Bennett
M R
MR
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000493:pharmacokinetics; D057846:Cell-Penetrating Peptides; D002478:Cells, Cultured; D005089:Exocytosis / Q000502:physiology; D005456:Fluorescent Dyes; D008565:Membrane Proteins / Q000737:chemistry* / Q000493:pharmacokinetics*; D008969:Molecular Sequence Data; D018377:Neurotransmitter Agents / Q000378:metabolism; D017729:Presynaptic Terminals / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011726:Pyridinium Compounds; D050765:Qa-SNARE Proteins; D000644:Quaternary Ammonium Compounds; D050683:R-SNARE Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D050600:SNARE Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D017783:Superior Cervical Ganglion / Q000166:cytology; D013572:Synaptic Vesicles / Q000378:metabolism*; D033921:Vesicular Transport Proteins
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; C073804:FM1 43; D005456:Fluorescent Dyes; D008565:Membrane Proteins; D018377:Neurotransmitter Agents; D011726:Pyridinium Compounds; D050765:Qa-SNARE Proteins; D000644:Quaternary Ammonium Compounds; D050683:R-SNARE Proteins; D050600:SNARE Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D033921:Vesicular Transport Proteins; C414409:penetratin