Thromb Haemost 1982; 48(02): 136-141
DOI: 10.1055/s-0038-1657242
Original Article
Schattauer GmbH Stuttgart

Effect of Pyridoxal 5'-Phosphate (PALP) on Human Platelet Aggregation, Dense Granule Release and Thromboxane B2 Generation - Role of Schiff Base Formation

S Krishnamurthi
The Thrombosis Research Unit, King's College Hospital Medical School, Denmark Hill, London, U.K.
,
J Westwick
The Thrombosis Research Unit, King's College Hospital Medical School, Denmark Hill, London, U.K.
,
V V Kakkar
The Thrombosis Research Unit, King's College Hospital Medical School, Denmark Hill, London, U.K.
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 25. März 1982

Accepted 12. Juli 1982

Publikationsdatum:
13. Juli 2018 (online)

Summary

Pyridoxal 5’-phosphate (PALP) inhibited ADP, thrombin, adrenaline, PAF and AA induced aggregation and 14C-5HT release. Thromboxane B2 (TxB2) generation induced by all the above agents except AA was also inhibited indicating that PALP may be inhibiting AA release via phospholipase A2 activation rather than AA metabolism. PALP inhibited ristocetin induced aggegation in PRP and agglutination in formaldehyde-treated washed platelets (FWP). Inhibition of ADP, adrenaline, PAF and AA-induced aggregation and 14C-5HT release by PALP was found in resuspended platelets pretreated with PALP and sodium borohydride suggesting that inhibition was mediated by Schiff base formation with platelet surface amino groups.

Irreversible fixation of PALP to the platelet membrane by borohydride reduction also inhibited thrombin induced 14C-5HT release and TxB2 generation but not thrombin induced primary aggregation or ristocetin induced agglutination in FWP. This suggests that PALP may interact with specific glycoproteins on the platelet membrane involved in ADP, adrenaline and PAF induced primary aggregation and that PALP could be inhibiting ristocetin induced agglutination by direct interaction with ristocetin or FVIII RCoF.

 
  • References

  • 1 Fasella P. Pyridoxal Phosphate. Ann Rev Biochem 1967; 36: 185-203
  • 2 Gold G, Widnell CC. Relationship between microsomal membrane permeability and the inhibition of hepatic glucose-6-phosphatase by pyridoxal phosphate. J Biol Chem 1976; 251: 1034-1041
  • 3 Wimmer MS, Sawyer DL, Harrison JH. Biphasic inactivation of porcine heart mitochondrial malate dehydrogenase by pyridoxal 5' phosphate. J Biol Chem 1975; 250: 710-715
  • 4 Colombo G, Marcus F. Modification of fructose-1,6 diphosphatase with pyridoxal 5' phosphate. Evidence for the participation of lysyl residues at the active site. Biochemistry 1974; 13: 3085-3091
  • 5 Raetz CRH, Auld DS. Schiff-base of pyridoxal phosphate with active centre lysins of ribonuclease A. Biochemistry 1972; 11: 2229-2236
  • 6 Eger R, Rifkin DB. The preparation and use of pyridoxal [32p]-phosphate as a labelling reagent for proteins on the outer surface of membranes. Biochim Biophys Acta 1977; 470: 70-83
  • 7 Cabantchik IZ, Balshin M, Breuer W, Rothstein A. Pyridoxal phosphate. An anionic probe for protein amino groups exposed on the outer and inner surfaces of intact human red blood cells. J Biol Chem 1975; 250: 5130-5136
  • 8 Subbarao K, Kuchibhotla J, Green D. Pyridoxine-induced inhibition of platelet aggregation and the release reaction. Circulation 1977; 56: 111-177
  • 9 Subbarao K, Kuchibhotla J, Kakkar VV. Pyridoxal 5' phosphate - a new physiological inhibitor of blood coagulation and platelet function. Biochem Pharmacol 1979; 28: 531-534
  • 10 Subbarao K, Kakkar VV, Ganguly P. Binding of pyridoxal phosphate to human platelets: its effect on platelet function. Thromb Res 1978; 13: 1017-1029
  • 11 Kornecki E, Feinberg H. Pyridoxal phosphate inhibition of platelet function. Am J Physiol 1980; 238: H54-H60
  • 12 Pollard HB, Tack-Goldman K, Pazoles CJ, Creutz CE, Shulman NR. Evidence for control of serotonin secretion from human platelets by hydroxyl ion transport and osmotic lysis. Proc Natl Acad Sd USA 1977; 74: 5295-5299
  • 13 Kornecki E, Feinberg H. Mechanism of inhibition of thrombin-induced platelet aggregation by pyridoxal phosphate. Biochem Biophys Res Commun 1979; 90: 963-968
  • 14 Demopoulos CA, Pinckard RN, Hanahan DJ. Platelet-activating factor. Evidence for 1-0-alkyl-2-acetyl-sn-glyceryl-3-phosphoryl choline as the active component. J Biol Chem 1979; 254: 9355-9358
  • 15 Bundy GL. The synthesis of prostaglandin endoperoxide analogues. Tetrahedron Lett 1975; 24: 1957
  • 16 Westwick J, Webb H. Selective antagonism of prostaglandin (PG) Elf PGD2 and prostacyclin (PGI2) on human and rabbit platelets by Di-4-phloretin phosphate (DPP). Thromb Res 1978; 12: 973-978
  • 17 Born GVR. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962; 194: 927-929
  • 18 Mills DCB, Robb IA, Roberts GCK. The release of nucleotides, 5-hydroxytryptamine and enzymes from human blood platelets during aggregation. J Physiol 1968; 195: 715-729
  • 19 Levy RI, Smith JB, silver MJ, Saia J, Walinsky P, Weiner L. Detection of thromboxane B2 in peripheral blood of patients with Prinzmetal’s angina. Prostaglandins Med 1979; 2: 243-244
  • 20 Lam SCT, Harfenist EJ, Packham MA, Mustard JF. Investigation of possible mechanism of pyridoxal 5' phosphate inhibition of platelet reactions. Thromb Res 1980; 20: 633-645
  • 21 Stoltz JF, Nicolas A. Study of amino groups of the human platelet membrane. Acta Haematol 1978; 60: 304-309
  • 22 Griffith MJ, Kingdon HS, Lundblad RL. The interaction of fibrinogen, antithrombin and heparin with pyridoxal phosphate modified human alpha-thrombin. Circ Res 1978; 57: 531
  • 23 Griffith MJ. Covalent modification of human α thrombin with pyridoxal 5' phosphate. J Biol Chem 1979; 254: 3401-3406
  • 24 Lapetina EG, Billah MM, Cuatrecasas P. The phosphatidyl inositol cycle and the regulation of arachidonic acid production. Nature 1981; 292: 367-369
  • 25 Dempsey WB, Chistensen HN. The specific binding of pyridoxal 5' phosphate to bovine plasma albumin. J Biol Chem 1962; 237: 1111-1120
  • 26 Stuart MJ, Gerrard JM, White JG. The influence of albumin and calcium on human platelet arachidonic acid metabolism. Blood 1980; 55: 418-423
  • 27 McDonald JWD, Stuart RK. Regulation of cyclic-AMP levels and aggregation in human platelets by prostaglandin E1 . J Lab Clin Med 1973; 81: 838-849
  • 28 Zahavi M, Kakkar VV. Adenosine 3' 5'-cyclic monophosphate (cAMP) and inhibitors of platelet aggregation: A possibility of a double cAMP pool in human platelets. Thromb Haemostas 1981; 46: 415 (Abstr)
  • 29 Coller BS. Inhibition of von Willebrand factor-dependent platelet function by increased platelet cyclic AMP and its prevention by cytoskeleton-disrupting agents. Blood 1981; 57: 846-855
  • 30 Weiss HJ, Rogers J, Brand H. Defective ristocetin-induced platelet aggregation in von Willebrand’s disease and its correction by factor VIII. J Clin Invest 1973; 52: 2697-2707
  • 31 Ganguly P. Binding of thrombin to functionally defective platelets: A hypothesis on the nature of the thrombin receptor. Br J Haematol 1977; 37: 47-51
  • 32 Jamieson GA, Okumura T. Platelet glycocalidn: A single receptor of platelet aggregation induced by thrombin or ristocetin. Thromb Res 1976; 8: 701-706
  • 33 Jamieson GA, Okumura T. Reduced thrombin binding and aggregation in Bernard-Soulier platelets. J Clin Invest 1978; 61: 861-864