Photolysis of a caged peptide reveals rapid action of N-ethylmaleimide sensitive factor before neurotransmitter release
- *Department of Neurobiology, Duke University Medical Center, Box 3209, Durham, NC 27710;
- ‡Marine Biological Laboratory, Woods Hole, MA 02543;
- §Molecular Probes Invitrogen, Eugene, OR 97402; and
- ¶School of Dentistry, University of Missouri, Kansas City, MO 64108-2784
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Edited by Richard W. Tsien, Stanford University School of Medicine, Stanford, CA, and approved November 21, 2007 (received for review August 1, 2007)
Abstract
The time at which the N-ethylmaleimide-sensitive factor (NSF) acts during synaptic vesicle (SV) trafficking was identified by time-controlled perturbation of NSF function with a photoactivatable inhibitory peptide. Photolysis of this caged peptide in the squid giant presynaptic terminal caused an abrupt (0.2 s) slowing of the kinetics of the postsynaptic current (PSC) and a more gradual (2–3 s) reduction in PSC amplitude. Based on the rapid rate of these inhibitory effects relative to the speed of SV recycling, we conclude that NSF functions in reactions that immediately precede neurotransmitter release. Our results indicate the locus of SNARE protein recycling in presynaptic terminals and reveal NSF as a potential target for rapid regulation of transmitter release.
Footnotes
- †To whom correspondence should be sent at present address: Department of Anatomy and Cell Biology, University of Heidelberg, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany. E-mail: kuner{at}uni-heidelberg.de
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Author contributions: T.K. and G.J.A. designed research; T.K. and Y.L. performed research; K.R.G. and L.F.B. contributed new reagents/analytic tools; T.K. and Y.L. analyzed data; and T.K. and G.J.A. wrote the paper.
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The authors declare no conflict of interest.
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This article is a PNAS Direct Submission.
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This article contains supporting information online at www.pnas.org/cgi/content/full/0707197105/DC1.
- © 2008 by The National Academy of Sciences of the USA
