Thromb Haemost 1993; 69(03): 286-292
DOI: 10.1055/s-0038-1651597
Original Article
Platelets
Schattauer GmbH Stuttgart

Trimucytin: A Collagen-Like Aggregating Inducer Isolated from Trimeresurus mucrosquamatus Snake Venom

Che-Ming Teng
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
,
Feng-Nien Ko
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
,
Inn-Ho Tsai
1   Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan
,
Man-Ling Hung
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
,
Tur-Fu Huang
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
› Author Affiliations
Further Information

Publication History

Received 29 January 1992

Accepted after revision 16 November 1992

Publication Date:
05 July 2018 (online)

Summary

Trimucytin is a potent platelet aggregation inducer isolated from Trimeresurus mucrosquamatus snake venom. Similar to collagen, trimucytin has a run of (Gly-Pro-X) repeats at the N-terminal amino acids sequence. It induced platelet aggregation, ATP release and thromboxane formation in rabbit platelets in a concentration-dependent manner. The aggregation was not due to released ADP since it was not suppressed by creatine phosphate/creatine phosphokinase. It was not either due to thromboxane A2 formation because indomethacin and BW755C did not have any effect on the aggregation even thromboxane B2 formation was completely abolished by indomethacin. Platelet-activating factor (PAF) was not involved in the aggregation since a PAF antagonist, kadsurenone, did not affect. However, RGD-containing peptide triflavin inhibited the aggregation, but not the release of ATP, of platelets induced by trimucytin. Indomethacin, mepacrine, prostaglandin E1 and tetracaine inhibited the thromboxane B2 formation of platelets caused by collagen and trimucytin. Forskolin and sodium nitroprusside inhibited both platelet aggregation and ATP release, but not the shape change induced by trimucytin. In quin-2 loaded platelets, the rise of intracellular calcium concentration caused by trimucytin was decreased by 12-O-tetradecanoyl phorbol-13 acetate, imipramine, TMB-8 and indomethacin. In the absence of extracellular calcium, both collagen and trimucytin caused no thromboxane B2 formation, but still induced ATP release which was completely blocked by R 59022. Inositol phosphate formation in platelets was markedly enhanced by trimucytin and collagen. MAB1988, an antibody against platelet membrane glycoprotein Ia, inhibited trimucytinand collagen-induced platelet aggregation and ATP release. However, trimucytin did not replace the binding of 125I-labeled MAB1988 to platelets. Platelets pre-exposed to trimucytin were resistant to the second challenge with trimucytin itself or collagen. It is concluded that trimucytin may activate collagen receptors on platelet membrane, and cause aggregation and release mainly through phospholipase C-phosphoinositide pathway.

 
  • References

  • 1 Teng CM, Huang TF. Snake venom constituents affect platelet function. Platelets 1991; 2: 1-11
  • 2 Brinkhous KM, Smith SV. Platelet-aggregating noncoagulant snake venom fractions. In: Hemostasis and Animal Venoms. Pirkle H, Markland Jr FS. (eds). New York: Marcel Dekker; 1988: 363-376
  • 3 Vargaftig BB. Platelet activation by non-coagulant snake venom components. Toxicon 1982; 20: 279-287
  • 4 Seegers WH, Ouyang C. Snake venoms and blood coagulation. In: Snake venoms, Handbook of Experimental Pharmacology. Lee CY. (ed) Berlin: Springer Verlag; 1979. 52 684-750
  • 5 Teng CM, Huang TF. Inventory of exogenous inhibitors of platelet aggregation. Thromb Haemostas 1991; 65: 624-626
  • 6 Chen JH, Wan H, Ran C. Activation of human platelets induced by the aggregoserpentin of Trimeresurus mucrosquamatus venom (TMVA). In: Hemostasis and Animal Venoms. Pirkle H, Markland Jr FS. (eds). New York: Marcel Dekker; 1988: 411-421
  • 7 Wal F, Joseph D, Tougui L, Vargaftig BB. PAF-acether may not mediate the third pathway of platelet aggregation since self-desensitization reduces the effects of low thrombin but enhances those of convulxin. Thomb Haemostas 1985; 53: 99-104
  • 8 Teng CM, Liao KK, Wang JP, Lin HS, Ouyang C. Ultrastructural changes and release reaction of platelets induced by an aggregation inducer purified from Trimeresurus mucrosquamatus (Formosan Habu) snake venom. Biochim Biophys Acta 1985; 841: 8-14
  • 9 Ouyang C, Wang JP, Teng CM. A potent platelet aggregation inducer purified from Trimeresurus mucrosquamatus snake venom. Biochim Biophys Acta 1980; 630: 246-253
  • 10 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . London: Nature 1970; 227: 680-685
  • 11 Hunkapiller MW. Users Manual, Applied Biosystems Model 120A PTH Analyzer. Applied Biosystems; Foster City, CA: 1986
  • 12 Prockop DJ. Mutations that alter the primary structure of type I collagen. J Biol Chem 1990; 265: 15349-15352
  • 13 Saus J, Quinones S, Mackrell AJ, Blumberg B, Muthuskumaran G, Pihlajaniemi T, Kurkinen M. The complete primary structure of mouse α2 (IV) collagen. J Biol Chem 1989; 264: 6318-6324
  • 14 Huang TF, Sheu JR, Teng CM, Chen SW, Liu CS. Triflavin, an antiplatelet Arg-Gly-Asp-containing peptide, is a specific antagonist of platelet membrane glycoprotein IIb/IIIa complex. J Biochem 1991; 109: 328-334
  • 15 Ardlie NG, Perry DW, Packham MA, Mustard JF. Influence of apyrase on stability of suspensions of washed rabbit platelets. Proc Soc Exp Biol Med 1971; 136: 1021-1023
  • 16 Born GVR, Cross MJ. The aggregation of blood platelets. J Physiol 1963; 168S: 178-195
  • 17 Teng CM, Ko FN. Comparison of the platelet aggregation induced by three thrombin-like enzymes of snake venoms and thrombin. Thromb Haemostas 1988; 59: 304-309
  • 18 DeLuca M, McElory WD. Purification and properties of firefly luciferase. Method Enzymol 1978; 57: 3-15
  • 19 Rink TJ, Sanchez A, Hallam TJ. Diacylglycerol and phorbol ester stimulate secretion without raising cytoplasmic free calcium in human platelets. Nature 1983; 305: 317-319
  • 20 Tsien RY, Pozzan T, Rink TJ. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol 1982; 94: 325-334
  • 21 Feinstein MB, Feiker J, Fraser C. An analysis of the mechanism of local anesthetics inhibition of platelet aggregation and secretion. J Pharmacol Exp Ther 1976; 197: 215-228
  • 22 Huang EM, Detwiler TC. The effect of lithium on platelet phosphoinositide metabolism. Biochem J 1986; 236: 895-901
  • 23 Neylon CB, Summers RJ. Stimulation of α1-adrenoceptors in rat kidney mediates increased inositol phospholipid hydrolysis. Br J Pharmacol 1987; 91: 367-376
  • 24 Higgs GA, Flower RJ, Vane JR. A new approach to anti-inflammatory drugs. Biochem Pharmacol 1979; 28: 1959-1961
  • 25 Packham MA, Guccione MA, Greenberg JP, Kinlough-Rathbone RL, Mustard JF. Release of 14C-serotonin during initial platelet change induced by thrombin, collagen or A23187. Blood 1977; 50: 915-926
  • 26 Vargaftig BB, Chignard M, Benveniste J. Present concepts on the mechanisms of platelet aggregation. Biochem Pharmacol 1981; 30: 263-277
  • 27 Braquet P, Godforid JJ. PAF-acether specific binding site: 2. Design of specific antagonist. Trends Pharmacol Sci 1986; 7: 297-403
  • 28 Rithenhouse SE, Allen CL. Synergistic activation by collagen and 15-hydroxy 9, 11-peroxido-prosta, 5, 13 dienoic acid (PGH2) of phosphatidylinositol metabolism and arachidonic acid release in human platelets. J Clin Invest 1982; 70: 1216-1224
  • 29 Haslam RJ, Lynham JA, Fox JEB. Effects of collagen, ionophore A23187 and prostaglandin E1 on the phosphorylation of specific proteins in blood platelets. Biochem J 1979; 178: 397-406
  • 30 Majerus PW, Wilson DB, Conolly TM, Bross TE, Neufeld EJ. Phosphoinositide turnover provides a link in stimulus-response coupling. Trends Biochem Sci 1985; 10: 168-71
  • 31 Knight DE, Scrutton MC. Direct evidence for a role for Ca2+ in amine storage granule secretion by human platelets. Thromb Res 1980; 20: 437-446
  • 32 Nishizuka Y. Phospholipid degradation and signal translation for protein phosphorylation. Trends Biochem Sci 1983; 8: 13-16
  • 33 Takai Y, Kaibuchi K, Mutsubara T, Nishizuka Y. Inhibitory action of guanosine 3' 5'-monophosphate on thrombin-induced phosphatidylinositol turnover and protein phosphorylation in human platelets. Biochem Biophys Res Commun 1981; 101: 61-67
  • 34 Hofmann SL, Prescott SM, Majerus PW. The effects of mepacrine and p-bromophenacyl bromide on arachidonic acid release in human platelets. Arch Biochem Biophys 1982; 215: 237-244
  • 35 Rittenhouse SE, Sasson JP. Mass changes in myoinositol triphosphate in human platelets stimulated by thrombin. J Biol Chem 1985; 260: 8657-8660
  • 36 de Chaffoy de Courcelles D, Roeven P, Vanbelle H. R 59022, a diacylglycerol kinase inhibitor: its effect on diacylglycerol and thrombin-induced C kinase activation in the intact platelet. J Biol Chem 1985; 260: 15762-15770
  • 37 Insel PA, Stengel D, Ferry N, Hanoune J. Regulation of adenyl cyclase of human platelet membranes by forskolin. J Biol Chem 1982; 257: 7485-7490
  • 38 Kawahara Y, Yamanishi J, Fukuzaki H. Inhibitory action of guanosine 3'5'-monophosphate on thrombin-induced calcium mobilization in human platelets. Thromb Res 1984; 33: 203-209
  • 39 Charo IF, Feinman RD, Detwiler TC. Inhibition of platelet secretion by an antagonist of intracellular calcium. Biochem Biophys Res Commun 1976; 72: 1462-1467
  • 40 Vanderhoek JY, Feinstein MB. Local anesthetics, chlorpromazine and propranolol inhibit stimulus activation of phospholipase, A2 in human platelets. Mol Pharmacol 1979; 16: 171-180
  • 41 Kunicki TJ, Nugent DJ, Staats SJ, Orchekowski RP, Wayner EA, Carter WG. The human fibroblast Class II extracellular matrix receptor mediates platelet adhesion to collagen is identical to the platelet glycoprotein Ia-IIa complex. J Biol Chem 1988; 263: 4516-4519
  • 42 Staatz WD, Rajpara SM, Wayner EA, Carter WG, Santoro SA. The membrane glycoprotein Ia-IIa (VLA-2) complex mediates the Mg2+- dependent adhesion of platelets to collagen. J Cell Biol 1989; 108: 1917-1924