A novel inhibitor of N-ethylmaleimide-sensitive factor decreases leukocyte trafficking and peritonitis.
Endothelial exocytosis is an early stage in the process of leukocyte trafficking. N-ethylmaleimide-sensitive factor (NSF) plays a critical role in regulating exocytosis. We hypothesized that inhibitors of NSF decrease endothelial exocytosis and vascular inflammation. We designed a novel fusion polypeptide consisting of a human immunodeficiency virus transactivator of transcription (TAT) protein transduction domain joined to a NSF homohexamerization domain. We show that this TAT-NSF polypeptide inhibits the ATPase activity and the disassembly activity of NSF. Furthermore, the TAT-NSF polypeptide decreases endothelial cell exocytosis and reduces leukocyte adherence to endothelial cells in culture. Finally, the TAT-NSF polypeptide inhibits leukocyte rolling on murine venules in vivo and inhibits leukocyte trafficking into the peritoneal cavity in a murine model of experimental peritonitis. These data suggest that NSF is a critical regulator of leukocyte trafficking in vivo. Novel compounds that inhibit the exocytic machinery in endothelial cells may be useful anti-inflammatory drugs.
Morrell
Craig N
CN
Department of Comparative Medicine, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Matsushita
Kenji
K
Lowenstein
Charles J
CJ
D000818:Animals; D042783:Endothelial Cells / Q000187:drug effects; D005089:Exocytosis / Q000187:drug effects; D036904:Leukocyte Rolling / Q000187:drug effects; D007962:Leukocytes / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D052066:N-Ethylmaleimide-Sensitive Proteins; D010529:Peritoneal Cavity / Q000166:cytology; D010538:Peritonitis / Q000188:drug therapy* / Q000473:pathology*; D016028:Receptors, Leukocyte-Adhesion / Q000187:drug effects; D050600:SNARE Proteins; D014699:Venules / Q000166:cytology; D033921:Vesicular Transport Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism
D016028:Receptors, Leukocyte-Adhesion; D050600:SNARE Proteins; D033921:Vesicular Transport Proteins; D052066:N-Ethylmaleimide-Sensitive Proteins; C498733:Nsf protein, mouse