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Membrane Cholesterol Influences the Activity of Recombinant Vibrio Vulnificus Hemolysin
Hui Ding1, Li-Ni Pan1, Qiu Guo1, Yong-Liang Lou1*, Jie Yan2,Yue Wu1 , Fu-Le Wang1
1Zhejiang Provincial Key Laboratory of Medical Genetics, School of Medical Lab Science , Wenzhou Medical College,Wenzhou 325035, China; 2Department of Medical Microbiology and Parasitology, College of Medicine, Zhejiang University, H
Abstract: To study the effect of membrane cholesterol on the activity of recombinant Vibrio vulnificus hemolysin (rVvhA). Before treated with rVvhA， J774A.1 cells were incubated with 5 mmol/L MβCD for 30 min to remove cholesterol from cells. After rVvhA treatment， the cell viability was determined by MTT assay. The intracellular [Ca2+] was determined using Fluo-3 AM fluorescent probe combined with Laser scanning confocal microscope， the release of cellular K+ into reaction medium and the cell apoptosis were detected according to detect kit instructions. The results showed cell pretreated with MβCD increased the cell viability， blocked rVvhA induced Ca2+ influx and K+ release， and inhibited the rVvhA induced-apoptosis. In conclusion， membrane cholesterol is required for activity of rVvhA， the depletion of cholesterol can inhibited the activity of rVvhA.
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