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Regulated Secretion in Platelets: Identification of Elements of the Platelet Exocytosis Machinery
Paula P. Lemons,
Dong Chen,
Audrey M. Bernstein,
Mark K. Bennett, and
S.W. Whiteheart
From the Department of Biochemistry, University of Kentucky College of Medicine, Lexington, KY; and the Department of Molecular and Cellular Biology, University of California, Berkeley CA.
To further characterize the molecular mechanisms of platelet function, we have sought to identify some of the proteins that mediate the secretory events of the platelet release reaction. We report that platelets contain the general elements of the membrane transport apparatus: N-ethylmaleimide sensitive fusion protein (NSF ), p115/transcytosis-associated protein (p115/TAP), and the soluble NSF attachment proteins ( - and,  -SNAP). The cDNAs encoding two of these proteins, - and -SNAP, have been cloned from a human platelet-derived cDNA library. Platelet membrane extracts possess SNAP receptor (SNARE) activity, suggesting that the class of proteins (SNAREs) proposed to provide the specificity for vesicle docking and membrane fusion are present in platelets. To identify these proteins, we have used specific antibodies against known SNAREs to probe platelet extracts. Syntaxin 2 and 4 can be readily detected in platelet membrane preparations and are shown to participate in 20 S complex formation. Syntaxin 1, 3, and 5 could not be detected. Other known SNARE and SNARE-associated proteins such as vesicle-associated membrane protein (VAMP)/synaptobrevin 2, SNAP-25, synaptophysin, or synaptotagmin I could not be immunochemically detected in platelet membrane preparations. The presence of both the general transport proteins (NSF and SNAPs) and specific transport proteins (syntaxin 2 and 4) indicates that platelet exocytosis uses a molecular mechanism similar to other secretory cells such as neurons. However, the subcellular concentrations of these proteins suggest that, unlike neuronal secretion, granule-to plasma membrane docking may be the limiting step in platelet exocytosis.
Blood, Vol. 90 No. 4 (August 15), 1997:
pp. 1490-1500
© 1997 by The American Society of Hematology.
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