Metabolic Properties of Human Platelet Membranes

Hermann Dietrich Kaulen; Rudolf Gross

doi:10.1055/s-0038-1649117

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1973; 30(01): 199-214
DOI: 10.1055/s-0038-1649117

Original Article

Schattauer GmbH

Metabolic Properties of Human Platelet Membranes

I. Characterization of Platelet Membranes prepared by Sucrose and Ficoll Density Gradients

Hermann Dietrich Kaulen

¹Medical Clinic, University of Cologne

,

Rudolf Gross

¹Medical Clinic, University of Cologne

› Author Affiliations

Abstract

Summary

The preparation of human platelet membranes and their characterization by electron microscopy and their enzymatic, lipid and protein content is reported. The membranes can be prepared in good yield from about one unit of blood. Using both sucrose and Ficoll density gradients as controls to each other, constant and reproducible membrane preparations of comparable biochemical composition are obtained. Electron microscopy shows uniform membrane structures without other contaminating subcellular organelles. The membranes are characterized by the enrichment of the acid para-nitrophenylphosphatase, a low activity of granular enzymes and by proteins of molecular weights from 20.000 to more than 100.000. They show a high content of N-acetylneuraminic acid, cholesterol, phospholipids, and in particular a high percentage of sphingomyelin. This preparation can be used for metabolic tracer studies at a subcellular level.

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References

References
1 Ashwoeth L. A. E, and Green C. 1966; Plasma Membranes: phospholipid and Sterol content. Science 151: 210.
2 Baggiolini M, Hirsch J. G, and de Duve Ch. 1970; Further Biochemical and Morphological Studies of Granule Fractions from Rabbit Heterophil Leucocytes. Journal of Cell Biology 45: 586.
3 Baenziger N. L, Brodie G. N, and Majerus Ph. W. 1971; A Thrombin-sensitive Protein of Human Platelet Membranes. Proceedings of the National Academy of Sciences of the United States of America 68: 240.
4 Barber A. J, and Jamieson G. A. 1970; Isolation and Characterization of Plasma Membranes from Human Blood Platelets. Journal of Biological Chemistry 245: 6357.
5 Barber A. J, Pepper D.S, and Jamieson G. A. 1971; A Comparison of Methods for Platelet Lysis and the Isolation of Platelet Membranes. Thrombosis et Diathesis Haemorrhagica 36: 38.
6 Behnke O. 1968; Electron microscopic Observations on the Membrane Systems of the Rat Blood Platelet. Anatomical Record 158: 121.
7 Bergmeyer H. U. (ed.) 1970. Methoden der enzymatischen Analyse. 2nd ed.. Verlag Chemie; Weinheim:
8 Booyse F. M, Hoveke Th. P, Kisieleski D, and Rafelson M. E. 1972; Mechanism and Control of Platelet-Platelet Interactions. II. Microvascular Research 4: 199.
9 Booyse F. M, and Rafelson M. E. 1967; In vitro Incorporation of Amino Acids in the Contractile Protein of Human Blood Platelets. Nature 215: 283.
10 Bowers W. E, Finkenstaedt J. T, and de Duve Ch. 1967; Lysosomes in Lymphoid Tissues. I. The Measurement of Hydrolytic Activities in Whole Homogenates. Journal of Cell Biology 32: 325.
11 Cheetham R. D, Keenan T. W, Nyquist S, and Morre D. J. 1969; Biochemical Comparisons of Endoplasmic Reticulum, Golgi Apparatus and Plasma Membrane-Rich Cell Fractions from Rat Liver in Relation to Cytomembrane Differentiation. Journal of Cell Biology 43: 21a.
12 Cohen Ph, Beoekman M. J, Veekley A, Lisman J. W.W, and Derksen A. 1971; Quantification of Human Platelet Inositides and the Influence of Ionic Environment on Their Incorporation of Orthophosphate-³²P. Journal of Clinical Investigation 50: 762.
13 Day H. J, Ang G. A. T, and Holmsen H. 1972; Platelet Release Reaction during Clotting of Native Human Platelet Rich Plasma. Proceedings of the Sociely of Experimental Biology and Medicine 139: 717.
14 Day H. J, Holmseist H, and Hovig T. 1969 Subcellular particles of Human Platelets. Scandinavian Journal of Haematology, Suppl. 7.
15 Dreisbach R. H. 1965; Submicrogram Determination of Inorganic Phosphate in Biological Material. Analytical Biochemistry 10: 169.
16 De Duve Ch. 1969. The Lysosome in retrospect. In: Dingle J. T, and Fell H. B. (eds.) Lysosomes in Biology and Pathology. North Holland Pubi. Co.; Amsterdam and London: 3.
17 Fairbanks G, Steck T. L, and Wallach D. F. H. 1971; Electrophoretic Analysis of the Major Polypeptides of the Human Erythrocyte Membrane. Biochemistry 10: 2606.
18 Gross R, and Schneider W. 1971. Energy metabolism. In: Johnson S. (ed.) The Circulating Platelet. Academic Press; Inc., New York and London: 123.
19 Henning R, Kaulen H. D, and Stoffel W. 1970; Isolation and Chemical Composition of the Lysosomal and the Plasma Membrane of the Rat Liver Cell. Hoppe-Seyler’s Zeitschrift für physiologische Chemie 351: 1191.
20 Holmsen H, and Day H. J. 1970; The Selectivity of the Thrombin-induced Platelet Release Reaction: Subcellular Localization of Released and Retained Constituents. Journal of Laboratory and Clinical Medicine 75: 840.
21 Holmsen H, Day H. J, and Pimentel M. A. 1969; Adenine Metabolism of Blood Platelets. V. Subcellular Localization of some related Enzymes. Biochemical et Biophysica Acta 186: 244.
22 Hovig T. 1968 The Ultrastructure of Blood Platelets in Normal and Abnormal States. Series Haematologica 1,2.
23 Ireland D. M. 1967; Effect of Thrombin on the Radioactive Nucleotides of Human Washed Platelets. Biochemical Journal 105: 857.
24 Johnson J. D, and Cornatzer W. E. 1969; Phospholipid Composition of Microchondria and Microsomes of Liver and Kidney in Adreanalectomized, Hypophysectomized and Diabetic Rats. Proceedings of the Society for Experimental Medicine and Biology 131: 474.
25 Kaulen H. D. 1972; The Separation of Phosphatidylserine and -inositol by One-dimensional Thin Layer Chromatography of Lipid Extracts. Analytical Biochemistry 45: 664.
26 Kaulen H. D, and Gross R. 1971; The Differentiation of Acid Phosphatases of Human Blood Platelets. Thrombosis et Diathesis Haemorrhagica 26: 353.
27 Kaulen H. D, Henning R, and Stoffel W. 1970; Comparison of some Enzymes of the Lysosomal and the Plasma Membrane of the Rat Liver Cell. Hoppe-Seyler’s Zeitschrift für physiologische Chemie 351: 1555.
28 Keenan T. W, and Morre D. J. 1970; Phospholipid Class and Fatty Acid Composition of Golgi Apparatus Isolated from Rat Liver and Comparison with other Cell Fractions. Biochemistry 9: 19.
29 Koerner J. F, and Sinsheimer R. L. 1957; A desoxyribonuclease of calf spleen. Journal of Biological Chemistry 228: 1049.
30 Lewis N, and Majerus Ph. W. 1969; Lipid Metabolism in Human Platelets. II. Journal of Clinical Investigation 48: 2114.
31 Lowry O. H, Rosebrough N, Farr J, and Randall R.J. 1951; Protein Measurement with the Folin Reagent. Journal of Biological Chemistry 193: 265.
32 Marcus A. J. 1969; a Platelet Function. New England Journal of Medicine 280: 1213 1778, 1330.
33 Marcus A. J, Ullman H. L, and Safier L. B. 1969; b Lipid Composition of Subcellular Particles of Human Blood Platelets. Journal of Lipid. Research 10: 108.
34 Marcus A. J, Zucker-Franklin D, Safier L. B, and Ullman H. L. 1966; Studies on Human Platelets Granules and Membranes. Journal of Clinical Investigation 45: 14.
35 Pepper D. S, and Jamieson G. A. 1969; Studies on Glycoproteins. III. Isolation of Sialyl-glycopeptides from Human Platelet Membranes. Biochemistry 8: 3362.
36 Pfleger R. C, Anderson N. G, and Snyder F. 1968; Lipid Class and Fatty Acid Composition of Rat Liver Plasma Membranes isolated by Zonal Centrifugation. Biochemistry 7: 2826.
37 Siegel A, Burri P. H, Weibel E. R, Bettex-Galland M, and Lüscher E. F. 1971; Density Gradient Centrifugation and Electron Microscopie Characterization of Subcellular Fractions from Human Blood Platelets. Thrombosis et Diathesis Haemorrhagica 25: 252.
38 Siegel A, and Lüscher E. F. 1967; Non-identity of α-granules of Human Blood Platelets with typical lysosomes. Nature 215: 745.
39 Stormorken H. 1969 The release reaction as secretion. Scandinavian Journal of Haematology, suppl. 9.
40 Wallach D. F. H, and Kamat V. B. 1964; Plasma and Cytoplasmic fragments from Ehrlich Ascites Carcinoma. Proceedings of the National Academy of Sciences of the United States of America, (Wash.) 52: 721.
41 Warren L. 1959; The Thiobarbituric Assay of Sialic Acids. Journal of Biological Chemistry 234: 1971.
42 Zlatkis A, ZAK B, and Boyle A. 1953; A new Method for the direct Determination of Serum Cholesterol. Journal of Laboratory and Clinical Médecine 41: 486.