抄録
The method of plasma membrane isolation using polylysine beads which was developed by Jacobson and Branton was applied to human platelets. The method is based on the electrostatic binding of positively charged polylysine beads and negatively charged cells, followed by disruption of the cells. The portion of the surface membrane attached on the beads remains bound there after disruption and is separated from other cellular components by washing. Purity of the platelet membrane preparation obtained by This method was estimated by enrichment of the 125I specific activity of the bead membrane preparation isolated from the platelets whose surface was iodinated with lactoperoxidase and H2O2. Marker enzymes, bis (p-nitrophenyl) phosphate and Na, K-ATPase for membranes, cytochrome oxidase for mitochondria and beta-glucuronidase for granules, were also assayed. These studies showed that the membrane preparation isolated on beads was satisfactorily pure as compared with the preparation isolated by widely used sucrose gradient centrifugation method. Analysis by polyacrylamide gel electrophoresis revealed that almost all species of the surface glycoproteins were present on the membrane preparation isolated with polylysine beads. The polylysine beads technique is a rapid, reproducible and efficient method for preparation of considerably pure platelet plasma membranes.
