Membrane cholesterol is required for activity of Vibrio vulnificus cytolysin.
Vibrio vulnificus cytolysin (VVC) forms a pore in the plasma membrane and induces cytolysis of various cells including erythrocytes, neutrophil and endothelial cells. The cytolytic activity of VVC is inhibited by exogenously added cholesterol, suggesting that membrane cholesterol might be required for VVC cytolytic activity. However, there is no direct evidence that membrane cholesterol is involved in VVC-induced cytolysis. Herein we demonstrate that membrane cholesterol is required for binding of VVC to the plasma membrane. Membrane cholesterol depletion with methyl-beta-cyclodextrin inhibited VVC-induced K(+) release, 2-deoxy glucose release and Ca(2+) influx, which are indicators of VVC pore formation. The cholesterol depletion-induced blockage of VVC cytolysis was due to the inhibition of VVC binding to membrane. These findings suggest that interaction with cholesterol is required for activity of VVC.
Yu
Hong-Nu
HN
Department of Biochemistry, Institute of Medical Science, Chonbuk National University Medical School, Jeonju, South Korea.
Lee
Young-Rae
YR
Park
Kwang-Hyun
KH
Rah
So-Young
SY
Noh
Eun-Mi
EM
Song
Eun-Kyung
EK
Han
Myung-Kwan
MK
Kim
Byeong-Soo
BS
Lee
Sung-Ho
SH
Kim
Jong-Suk
JS
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D004912:Erythrocytes / Q000502:physiology; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D009504:Neutrophils / Q000382:microbiology; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins / Q000378:metabolism*; D051381:Rats; D041261:Vibrio vulnificus / Q000378:metabolism* / Q000472:pathogenicity*; D047392:beta-Cyclodextrins / Q000494:pharmacology
D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D054353:Perforin; D002784:Cholesterol