Abstract
Human platelet plasma membranes were isolated with polylysine beads according to the technique developed by Jacobson and Branton (1977, Science [Wash. D. C.] 195:302--304). Lactoperoxidase-catalyzed surface iodination revealed that ninefold greater 125I specific activity was associated with the membranes isolated on beads than with whole platelets. Enrichment in the bead membrane preparation of the activities of membrane marker enzymes, bis(p-nitrophenyl)phosphate phosphodiesterase and Na,K-ATPase, was 8.0 and 4.4, respectively. Contamination with enzymes of other organelles, cytochrome oxidase and beta-glucuronidase, was relatively low as compared with membranes isolated by sucrose gradient centrifugation. Analysis by SDS polyacrylamide gel electrophoresis showed that a full complement of surface glycoproteins was present on the membranes isolated with polylysine beads. The polylysine bead technique is a rapid, reproducible and efficient method for the preparation of relatively pure platelet plasma membranes.
Full Text
The Full Text of this article is available as a PDF (907.4 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Andersson L. C., Gahmberg C. G. Surface glycoproteins of human white blood cells. Analysis by surface labeling. Blood. 1978 Jul;52(1):57–67. [PubMed] [Google Scholar]
- Barber A. J., Jamieson G. A. Isolation and characterization of plasma membranes from human blood platelets. J Biol Chem. 1970 Dec 10;245(23):6357–6365. [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Broekman M. J., Westmoreland N. P., Cohen P. An improved method for isolating alpha granules and mitochondria from human platelets. J Cell Biol. 1974 Feb;60(2):507–519. doi: 10.1083/jcb.60.2.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen C. M., Kalish D. I., Jacobson B. S., Branton D. Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells. J Cell Biol. 1977 Oct;75(1):119–134. doi: 10.1083/jcb.75.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gahmberg C. G., Hakomori S. I. External labeling of cell surface galactose and galactosamine in glycolipid and glycoprotein of human erythrocytes. J Biol Chem. 1973 Jun 25;248(12):4311–4317. [PubMed] [Google Scholar]
- Guccione M. A., Packham M. A., Kinlough-Rathbone R. L., Perry D. W., Mustard J. F. Reactions of polylysine with human platelets in plasma and in suspensions of washed platelets. Thromb Haemost. 1976 Nov 30;36(2):360–375. [PubMed] [Google Scholar]
- Hatton M. W., Regoeczi E. A simple method for the purification of commercial neuraminidase preparations free from proteases. Biochim Biophys Acta. 1973 Nov 15;327(1):114–120. doi: 10.1016/0005-2744(73)90108-3. [DOI] [PubMed] [Google Scholar]
- Jacobson B. S., Branton D. Plasma membrane: rapid isolation and exposure of the cytoplasmic surface by use of positively charged beads. Science. 1977 Jan 21;195(4275):302–304. doi: 10.1126/science.831278. [DOI] [PubMed] [Google Scholar]
- Kalish D. I., Cohen C. M., Jacobson B. S., Branton D. Membrane isolation on polylysine-coated glass beads. Asymmetry of bound membrane. Biochim Biophys Acta. 1978 Jan 4;506(1):97–110. doi: 10.1016/0005-2736(78)90437-6. [DOI] [PubMed] [Google Scholar]
- Kaplan K. L., Nachman R. L. The effect of platelet membrane antibodies on aggregation and release. Br J Haematol. 1974 Dec;28(4):551–560. doi: 10.1111/j.1365-2141.1974.tb06674.x. [DOI] [PubMed] [Google Scholar]
- Marcus A. J., Zucker-Franklin D., Safier L. B., Ullman H. L. Studies on human platelet granules and membranes. J Clin Invest. 1966 Jan;45(1):14–28. doi: 10.1172/JCI105318. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Massini P., Metcalf L. C., Näf U., Lüscher E. F. Induction of aggregation and of the release reaction in human platelets by polylysine. Haemostasis. 1974;3(1):8–19. doi: 10.1159/000214038. [DOI] [PubMed] [Google Scholar]
- Nachman R. L., Ferris B. Binding of adenosine diphosphate by isolated membranes from human platelets. J Biol Chem. 1974 Feb 10;249(3):704–710. [PubMed] [Google Scholar]
- Nachman R. L., Hubbard A., Ferris B. Iodination of the human platelet membrane. Studies of the major surface glycoprotein. J Biol Chem. 1973 Apr 25;248(8):2928–2936. [PubMed] [Google Scholar]
- Nachman R. L., Tarasov E., Weksler B. B., Ferris B. Wheat germ agglutinin affinity chromatography of human platelet membrane glycoproteins. Thromb Res. 1978 Jan;12(1):91–104. doi: 10.1016/0049-3848(78)90088-9. [DOI] [PubMed] [Google Scholar]
- Newman K. D., Williams L. T., Bishopric N. H., Lefkowitz R. J. Identification of alpha-adrenergic receptors in human platelets by [3H]dihydroergocryptine binding. J Clin Invest. 1978 Feb;61(2):395–402. doi: 10.1172/JCI108950. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nurden A. T., Caen J. P. An abnormal platelet glycoprotein pattern in three cases of Glanzmann's thrombasthenia. Br J Haematol. 1974 Oct;28(2):253–260. doi: 10.1111/j.1365-2141.1974.tb06660.x. [DOI] [PubMed] [Google Scholar]
- Nurden A. T., Caen J. P. Specific roles for platelet surface glycoproteins in platelet function. Nature. 1975 Jun 26;255(5511):720–722. doi: 10.1038/255720a0. [DOI] [PubMed] [Google Scholar]
- Okumura T., Jamieson G. A. Platelet glycocalicin. I. Orientation of glycoproteins of the human platelet surface. J Biol Chem. 1976 Oct 10;251(19):5944–5949. [PubMed] [Google Scholar]
- Phillips D. R., Agin P. P. Platelet plasma membrane glycoproteins. Evidence for the presence of nonequivalent disulfide bonds using nonreduced-reduced two-dimensional gel electrophoresis. J Biol Chem. 1977 Mar 25;252(6):2121–2126. [PubMed] [Google Scholar]
- Phillips D. R., Agin P. P. Platelet plasma membrane glycoproteins. Identification of a proteolytic substrate for thrombin. Biochem Biophys Res Commun. 1977 Apr 25;75(4):940–947. doi: 10.1016/0006-291x(77)91473-5. [DOI] [PubMed] [Google Scholar]
- Phillips D. R. Effect of trypsin on the exposed polypeptides and glycoproteins in the human platelet membrane. Biochemistry. 1972 Nov 21;11(24):4582–4588. doi: 10.1021/bi00774a025. [DOI] [PubMed] [Google Scholar]
- Rinderknecht H., Geokas M. C., Silverman P., Haverback B. J. A new ultrasensitive method for the determination of proteolytic activity. Clin Chim Acta. 1968 Aug;21(2):197–203. doi: 10.1016/0009-8981(68)90127-7. [DOI] [PubMed] [Google Scholar]
- Sixma J. J., Lips P. M. Isolation of platelet membranes. A review. Thromb Haemost. 1978 Apr 30;39(2):328–337. [PubMed] [Google Scholar]
- Taylor D. G., Crawford N. The separation of platelet membranes and the various granular organelles by a simple density gradient procedure using the B14 zonal rotor. Biochem Med. 1975 Nov;14(3):324–338. doi: 10.1016/0006-2944(75)90051-4. [DOI] [PubMed] [Google Scholar]
- Taylor D. G., Crawford N. The subfractionation of platelet membranes by zonal centrifugation: identification of surface membranes. FEBS Lett. 1974 May 1;41(2):317–322. doi: 10.1016/0014-5793(74)81238-x. [DOI] [PubMed] [Google Scholar]
- Tollefsen D. M., Feagler J. R., Majerus P. W. The binding of thrombin to the surface of human platelets. J Biol Chem. 1974 Apr 25;249(8):2646–2651. [PubMed] [Google Scholar]
