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Thromb Haemost 1992; 67(05): 567-571
DOI: 10.1055/s-0038-1648495
Original Articles
Schattauer GmbH Stuttgart

Membrane Fluidity Is Related to the Extent of Glycation of Proteins, but not to Alterations in the Cholesterol to Phospholipid Molar Ratio in Isolated Platelet Membranes from Diabetic and Control Subjects

Peter D Winocour
The Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
Cezary Watala
The Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
Raelene L Kinglough-Rathbone
The Department of Pathology, McMaster University, Hamilton, Ontario, Canada
› Author Affiliations
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Publication History

Received 19 April 1991

Accepted after revision 20 November 1991

Publication Date:
03 July 2018 (online)

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Summary

Platelets from diabetic subjects are hypersensitive to aggregating agents in vitro. Membrane fluidity modulates cell function and we previously reported reduced membrane fluidity associated with hypersensitivity to thrombin in intact platelets from diabetic subjects. Reduced membrane fluidity and hypersensitivity to agonists has also been reported in platelets from non-diabetic subjects whose platelets have an increased cholesterol/phos-pholipid molar ratio. Glycation of platelet membrane proteins is enhanced in diabetic subjects, and could contribute to the decreased membrane fluidity in these platelets. We examined the relation among fluidity, cholesterol/phospholipid molar ratio, and glycation of proteins in isolated platelet membranes from diabetic and control subjects. Seven poorly controlled diabetic subjects were compared with 7 age- and sex-matched control subjects. The mean steady-state fluorescence polarization value in 1,6-diphenyl-1,3,5-hexatriene-labeled isolated platelet membranes from diabetic subjects (0.184 ± 0.004) was significantly greater than from control subjects (0.171 ± 0.004, p <0.01); thus, fluidity in platelet membranes from diabetic subjects is decreased. Reduced fluidity in platelet membranes from diabetic subjects could not be attributed to changes in the cholesterol/phospholipid molar ratio. Total or very low density (VLDL), low density (LDL), or high density (HDL3) lipoprotein cholesterol concentration in plasma was not significantly different between groups, but the ratio of VLDL + LDL to HDL2 + HDL3 cholesterol was significantly greater in diabetic subjects (4.79 ± 0.73) than in control subjects (2.54 ± 0.30, p <0.02). Proteins were glycated significantly more extensively in platelet membranes from diabetic subjects (25.5 ± 0.9 nmol glucose/mg protein) than those from control subjects (21.0 ± 0.6 nmol glucose/mg protein, p <0.001). The extent of glycation of proteins in platelet membranes correlated significantly with the steady state fluorescence polarization values for platelet membranes from diabetic and control subjects (r = 0.715, p <0.01). More extensive glycation of proteins appears to be associated with reduced fluidity of platelet membranes from diabetic subjects, but not with increased cholesterol/phospholipid molar ratio.

 

  • References

  • 1 Winocour PD. The role of platelets in the pathogenesis of diabetic vascular disease. In: Complications of Diabetes Mellitus. Molecular and Cellular Biology of Diabetes Mellitus, Vol III. Draznin B, Melmed S, LeRoith D. (eds) Alan R. Liss, Inc.; New York: 1989. pp 37-47
    Google Scholar
  • 2 Colwell JA, Winocour PD, Lopes-Virella MF. Platelet function and platelet-plasma interactions in atherosclerosis and diabetes mellitus. In: Diabetes Mellitus, Theory and Practice, 4th edition. Rifkin H, Porte D. (eds) Elsevier; New York: 1990. pp 249-256
    Google Scholar
  • 3 Colwell JA, Lopes-Virella MF, Winocour PD, Halushka PV. New concepts about the pathogenesis of atherosclerosis in diabetes mellitus. In: The Diabetic Foot. Levin ME, O’Neal LW. (eds) Mosby, St. Louis, MO: 1988. pp 51-70
    Google Scholar
  • 4 Banga JD, Sixma JJ. Diabetes mellitus, vascular disease and thrombosis. Clin Haematol 1986; 15: 465-492
    PubMedGoogle Scholar
  • 5 Ruderman NB, Haudenschild C. Diabetes as an atherogenic factor. Prog Cardiovasc Dis 1984; 26: 373-412
    CrossrefPubMedGoogle Scholar
  • 6 Shinitzky M, Henkart P. Fluidity of cell membranes: current concepts and trends. Int Rev Cytol 1979; 60: 121-147
    PubMedGoogle Scholar
  • 7 Tandon NN, Harmon JT, Jamieson GA. Influence of membrane fluidity on platelet activation in cholesterol-modified and hypercholes-terolemic platelets. In: Lipid Domains and the Relationship to Membrane Function. Aloia RC, Curtain CC, Gordon LM. (eds) Alan R. Liss Inc.; New York: 1988. pp 83-99
    Google Scholar
  • 8 Winocour PD, Bryszewska M, Watala C, Rand ML, Epand RM, Kinlough-Rathbone RL, Packham MA, Mustard JF. Reduced membrane fluidity in platelets from diabetic patients. Diabetes 1990; 39: 241-244
    CrossrefPubMedGoogle Scholar
  • 9 Shattil SJ, Cooper RA. Membrane microviscosity and human platelet function. Biochemistry 1976; 15: 4832-4837
    CrossrefPubMedGoogle Scholar
  • 10 Tandon N, Harmon JT, Rodbard D, Jamieson GA. Thrombin receptors define responsiveness of cholesterol-modified platelets. J Biol Chem 1983; 258: 11840-11845
    CrossrefPubMedGoogle Scholar
  • 11 Shattil SJ, Cooper RA. Role of membrane lipid composition, organization, and fluidity in human platelet function. Prog Hemostas Thromb 1978; 4: 59-86
    PubMedGoogle Scholar
  • 12 Shattil SJ, Bennett JS, Colman RW, Cooper RA. Abnormalities of cholesterol-phospholipid composition in platelets and low-density lipoproteins of human hyperbetalipoproteinemia. J Lab Clin Med 1977; 89: 341-353
    PubMedGoogle Scholar
  • 13 Harmon JT, Tandon NN, Hoeg JM, Jamieson GA. Thrombin binding and response in platelets from patients with dyslipoproteinemias: increased stimulus-response coupling in type II hyperlipoproteinemia. Blood 1986; 68: 498-505
    PubMedGoogle Scholar
  • 14 Watala C, Zawodniak M, Bryszewska M, Nowak S. Nonenzymatic protein glycosylation. I. Lowered erythrocyte membrane fluidity in juvenile diabetes. Ann Clin Res 1985; 17: 327-330
    PubMedGoogle Scholar
  • 15 Bryszewska M, Szosland K. Association between the glycation of erythrocyte membrane proteins and membrane fluidity. Clin Biochem 1988; 21: 49-51
    CrossrefPubMedGoogle Scholar
  • 16 Watala C. In vitro glycation of red blood cell proteins: high levels of glucose lower lipid fluidity of erythrocyte membranes. Exp Pathol 1988; 33: 233-238
    CrossrefPubMedGoogle Scholar
  • 17 Sampietro T, Lenzi S, Cecchetti P, Giampietro O, Cruschelli L, Navalesi R. Nonenzymatic glycation of human platelet membrane proteins in vitro and in vivo. Clin Chem 1986; 32: 1328-1331
    PubMedGoogle Scholar
  • 18 Yatscoff RW, Mehta A, Gerrard JM, Thliveris J. Glycation of platelet protein in diabetes mellitus: lack of correlation with platelet function. Clin Biochem 1987; 20: 359-363
    CrossrefPubMedGoogle Scholar
  • 19 Nair G, Sainani GS, Krishnaswamy PR. Evidence of glycosylation of platelet proteins in diabetics. J Assoc Phys India 1985; 33: 299-300
    PubMedGoogle Scholar
  • 20 Cohen I, Burk D, Fullerton RJ, Veis A, Green D. Nonenzymatic glycation of human blood platelet proteins. Thromb Res 1989; 55: 341-349
    CrossrefPubMedGoogle Scholar
  • 21 Aster RH, Jandl JH. Platelet sequestration in man. I. Methods. J Clin Invest 1964; 43: 843-855
    CrossrefPubMedGoogle Scholar
  • 22 Fox JEB, Say AK, Haslam RJ. Subcellular distribution of the different platelet proteins phosphorylated on exposure of intact platelets to ionophore A23187 or to prostaglandin Ex. Possible role of a membrane phosphopolypeptide in the regulation of calcium-ion transport. Biochem J 1979; 184: 651-661
    CrossrefPubMedGoogle Scholar
  • 23 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-275
    PubMedGoogle Scholar
  • 24 Shinitzky M, Barenholz Y. Fluidity parameters of lipid regions determined by fluorescence polarization. Biochim Biophys Acta 1978; 515: 367-394
    CrossrefPubMedGoogle Scholar
  • 25 Chen RF, Bowman RL. Fluorescence polarization - measurement with ultra-polarizing filters in a spectrophotofluorometer. Science 1965; 147: 729-732
    CrossrefPubMedGoogle Scholar
  • 26 Azumi T, McGlynn SP. Polarization of the luminescence of phenan-threne. J Chem Phys 1962; 37: 2413-2420
    CrossrefPubMedGoogle Scholar
  • 27 Van Blitterswijk WJ, van der Meer BW, Hilkmann H. Quantitative contributions of cholesterol and the individual classes of phospholipids and their degree of fatty acyl (un)saturation to membrane fluidity measured by fluorescence polarization. Biochemistry 1987; 26: 1746-1756
    CrossrefPubMedGoogle Scholar
  • 28 Berlin E, Shapiro SG, Friedland M. Platelet membrane fluidity and aggregation of rabbit platelets. Atherosclerosis 1984; 51: 223-239
    CrossrefPubMedGoogle Scholar
  • 29 Vickers JD, Rathbone MP. The effect of membrane cholesterol depletion upon erythrocyte membrane-bound proteins. Can J Biochem 1979; 57: 1144-1152
    CrossrefPubMedGoogle Scholar
  • 30 Gamble W, Vaughan M, Kruth HS, Avigan J. Procedure for determination of free and total cholesterol in micro- and nanogram amounts suitable for studies with cultured cells. J Lipid Res 1978; 19: 1068-1070
    PubMedGoogle Scholar
  • 31 Ames BN. Assay of inorganic phosphate, total phosphate and phosphates. In: Complex Carbohydrate. Neufeld EF, Ginsberg V. (eds) In: >Methods in Enzymology, Vol.VIII. Colowick SP, Kaplan NO. (eds.-in-chief) Academic Press; New York: 1966. pp 115-118
    Google Scholar
  • 32 Witztum JL, Mahoney EM, Branks MJ, Fisher M, Elam R, Steinberg D. Nonenzymatic glucosylation of low-density lipoprotein alters its biologic activity. Diabetes 1982; 31: 283-291
    CrossrefPubMedGoogle Scholar
  • 33 Allen DW, Schroeder WA, Balog J. Observation on chromatographic heterogeneity of normal adult and fetal haemoglobin. . Chem Soc Rev 1958; 80: 1628-1634
    PubMedGoogle Scholar
  • 34 Kadish AH, Little RL, Sternberg JC. A new and rapid method for the determination of glucose by measurement of rate of oxygen consumption. Clin Chem 1968; 14: 116-131
    PubMedGoogle Scholar
  • 35 Hatch FT, Lees RS. Practical methods for plasma lipoprotein analysis. Adv Lipid Res 1968; 6: 1-67
    PubMedGoogle Scholar
  • 36 Heemskerk JWM, Feijge MAH, Kalafusz R, Hornstra G. Influence of dietary fatty acids on membrane fluidity and activation of rat platelets. Biochim Biophys Acta 1989; 1004: 252-260
    CrossrefPubMedGoogle Scholar
  • 37 Berlin E, Matusik EJ, Young C. Effect of dietary fat on the fluidity of platelet membranes. Lipids 1980; 15: 604-608
    CrossrefPubMedGoogle Scholar
  • 38 Gibney MJ, Bolton-Smith C. The effect of a dietary supplement of n-3 plasma membrane fluidity in healthy volunteers. Br J Nutr 1988; 60: 5-12
    CrossrefPubMedGoogle Scholar
  • 39 Rand ML, Hennissen AAHM, Hornstra G. Effects of dietary sunflowerseed oil and marine oil on platelet membrane fluidity, arterial thrombosis, and platelet responses in rats. Atherosclerosis 1986; 62: 267-276
    CrossrefPubMedGoogle Scholar
  • 40 Steiner M. Vitamin E changes the membrane fluidity of human platelets. Biochim Biophys Acta 1981; 640: 100-105
    CrossrefPubMedGoogle Scholar
  • 41 Prisco D, Rogasi PG, Paniccia R, Abbate R, Gensini GF, Pinto S, Vanni D, Neri Serneri GG. Altered membrane fatty acid composition and increased thromboxane A2 generation in platelets from patients with diabetes. Prost Leuk Ess Fatty Acids 1989; 35: 15-23
    CrossrefPubMedGoogle Scholar
  • 42 Jones DB, Carter RD, Haitas B, Mann JI. Low phospholipid arachidonic acid values in diabetic platelets. Br Med J 1983; 286: 173-175
    CrossrefPubMedGoogle Scholar
  • 43 Karpen CW, Pritchard KA, Arnold JH, Cornwell DG, Panganamala RV. Restoration of prostacyclin/thromboxane A2 balance in the diabetic rat. Influence of dietary vitamin E. Diabetes 1982; 31: 947-951
    CrossrefPubMedGoogle Scholar
  • 44 Karpen CW, Cataland S, O’Dorisio TM, Panaganamala RV. Interrelation of platelet vitamin E and thromboxane synthesis in type I diabetes mellitus. Diabetes 1984; 33: 239-243
    CrossrefPubMedGoogle Scholar
  • 45 DiMinno G, Silver MJ, Cerbone AM, Riccardi G, Rivellese A, Mancini M. Platelet fibrinogen binding in diabetes mellitus. Differences between binding to platelets from non-retinopathic and retinopathic diabetic patients. Diabetes 1986; 35: 182-185
    CrossrefPubMedGoogle Scholar