Triplatin, a platelet aggregation inhibitor from the salivary gland of the triatomine vector of Chagas disease, binds to TXA2 but does notinteract with glycoprotein PVI

Dongying Ma; Teresa C. F. Assumpção; Yuan Li; John F. Andersen; José M.C. Ribeiro; Ivo M.B. Francischetti

doi:10.1160/TH11-10-0685

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2012; 107(01): 111-123
DOI: 10.1160/TH11-10-0685

Platelets and Blood Cells

Schattauer GmbH

Triplatin, a platelet aggregation inhibitor from the salivary gland of the triatomine vector of Chagas disease, binds to TXA₂ but does notinteract with glycoprotein PVI

Dongying Ma

¹Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

,

Teresa C. F. Assumpção

¹Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

,

Yuan Li

¹Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

,

John F. Andersen

¹Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

,

José M.C. Ribeiro

¹Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

,

Ivo M.B. Francischetti

¹Section of Vector Biology, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

› Author Affiliations

Abstract

Summary

Salivary glands from haematophagous animals express a notable diversity of negative modulators of platelet function. Triplatin is an inhibitor of collagen-induced platelet aggregation which has been described as an antagonist of glycoprotein VI (GPVI). Because triplatin displays sequence homology to members of the lipocalin family of proteins, we investigated whether triplatin mechanism of action could be explained by interaction with pro-haemostatic prostaglandins. Our results demonstrate that triplatin inhibits platelet aggregation induced by low doses of collagen, thromboxane A2 (TXA₂) mimetic (U46619), and arachidonic acid (AA). On the other hand, it does not inhibit platelet aggregation by convulxin, PMA, or low-dose ADP. Isothermal titration calorimetry (ITC) revealed that triplatin binds AA, cTXA₂, TXB₂, U46619 or prostaglandin (PG)H₂ mimetic (U51605). Consistent with its ligand specificity, triplatin induces relaxation of rat aorta contracted with U46619. Triplatin also interacts with PGF_2a and PGJ₂, but not with leukotrienes, AA or biogenic amines. Surface plasmon resonance experiments failed to demonstrate interaction of triplatin with GPVI; it also did to inhibit platelet adhesion to fibrillar or soluble collagen. Because triplatin displays sequence similarity to apolipoprotein D (ApoD) – a lipocalin associated with high-density lipoprotein, ApoD was tested as a putative TXA₂-binding molecule. ITC failed to demonstrate binding of ApoD to all prostanoids described above, or to AA. Furthermore, ApoD was devoid of inhibitory properties towards platelets activation by AA, collagen, or U46619. In conclusion, triplatin mechanism of action has been elucidated without ambiguity as a novel TXA₂- and PGF_2α- binding protein. It conceivably blocks platelet aggregation and vasoconstriction, thus contributing to successful blood feeding at the vector-host interface.

Keywords

Snake venoms - protein function / activity - platelet physiology

Full Text

References

References
1 Watson SP. Platelet activation by extracellular matrix proteins in haemostasis and thrombosis. Curr Pharm Des 2009; 15: 1358-1372.
2 Nieswandt B, Watson SP. Platelet-collagen interaction: is GPVI the central receptor?. Blood 2003; 102: 449-461.
3 Jung SM, Moroi M. Activation of the platelet collagen receptor integrin alpha(2)beta(1): its mechanism and participation in the physiological functions of platelets. Trends Cardiovasc Med 2000; 10: 285-292.
4 Cosemans JM, Iserbyt BF, Deckmyn H. et al. Multiple ways to switch platelet integrins on and off. J Thromb Haemost 2008; 06: 1253-1261.
5 Gibbins JM. Platelet adhesion signalling and the regulation of thrombus formation. J Cell Sci 2004; 117: 3415-3425.
6 Jackson SP. The growing complexity of platelet aggregation. Blood 2007; 109: 5087-5095.
7 Furie B, Furie BC. Mechanisms of thrombus formation. N Engl J Med 2008; 359: 938-949.
8 Giannarelli C, Zafar MU, Badimon JJ. Prostanoid and TP-receptors in athero-thrombosis: is there a role for their antagonism?. Thromb Haemost 2010; 104: 949-954.
9 Jaipersad AS, Silverman SH, Lip GY. Anti-platelet therapy: is it all over in peripheral artery disease?. Thromb Haemost 2010; 103: 689-691.
10 Ribeiro JM, Francischetti IM. Role of arthropod saliva in blood feeding: sialome and post-sialome perspectives. Annu Rev Entomol 2003; 48: 73-88.
11 Fry BG, Roelants K, Champagne DE. et al. The toxicogenomic multiverse: convergent recruitment of proteins into animal venoms. Annu Rev Genomics Hum Genet 2009; 10: 483-511.
12 Francischetti IM. Platelet aggregation inhibitors from hematophagous animals. Toxicon 2010; 56: 1130-1144.
13 Andersen JF. Structure and mechanism in salivary proteins from blood-feeding arthropods. Toxicon 2010; 56: 1120-1129.
14 Maritz-Olivier C, Stutzer C, Jongejan F. et al. Tick anti-hemostatics: targets for future vaccines and therapeutics. Trends Parasitol 2007; 23: 397-407.
15 Steen NA, Barker SC, Alewood PF. Proteins in the saliva of the Ixodida (ticks): pharmacological features and biological significance. Toxicon 2006; 47: 1-20.
16 Noeske-Jungblut C, Kratzschmar J, Haendler B. et al. An inhibitor of collagen-induced platelet aggregation from the saliva of Triatoma pallidipennis. J Biol Chem 1994; 269: 5050-5053.
17 Assumpcao TC, Alvarenga PH, Ribeiro JM. et al. Dipetalodipin, a novel multifunctional salivary lipocalin that inhibits platelet aggregation, vasoconstriction, and angiogenesis through unique binding specificity for TXA2, PGF2alpha, and 15(S)-HETE. J Biol Chem 2010; 285: 39001-39012.
18 Waxman L, Connolly TM. Isolation of an inhibitor selective for collagen-stimulated platelet aggregation from the soft tick Ornithodoros moubata. J Biol Chem 1993; 268: 5445-5449.
19 Mans BJ, Ribeiro JM. Function, mechanism and evolution of the moubatin-clade of soft tick lipocalins. Insect Biochem Mol Biol 2008; 38: 841-852.
20 Reddy VB, Kounga K, Mariano F. et al. Chrysoptin is a potent glycoprotein Ilb/IIIa fibrinogen receptor antagonist present in salivary gland extracts of the deerfly. J Biol Chem 2000; 275: 15861-15867.
21 Morita A, Isawa H, Orito Y. et al. Identification and characterization of a collagen-induced platelet aggregation inhibitor, triplatin, from salivary glands of the assassin bug, Triatoma infestans. FEBS J 2006; 273: 2955-2962.
22 Nofer JR, Brodde MF, Kehrel BE. High-density lipoproteins, platelets and the pathogenesis of atherosclerosis. Clin Exp Pharmacol Physiol 2010; 37: 726-735.
23 Francischetti IM, Saliou B, Leduc M. et al. Convulxin, a potent platelet-aggregating protein from Crotalus durissus terrificus venom, specifically binds to platelets. Toxicon 1997; 35: 1217-1228.
24 Jandrot-Perrus M, Busfield S, Lagrue AH. et al. Cloning, characterization, and functional studies of human and mouse glycoprotein VI: a platelet-specific collagen receptor from the immunoglobulin superfamily. Blood 2000; 96: 1798-1807.
25 Francischetti IM, Ribeiro JM, Champagne D. et al. Purification, cloning, expression, and mechanism of action of a novel platelet aggregation inhibitor from the salivary gland of the blood-sucking bug, Rhodnius prolixus. J Biol Chem 2000; 275: 12639-12650.
26 Paddock CD, McKerrow JH, Hansell E. et al. Identification, cloning, and recombinant expression of procalin, a major triatomine allergen. J Immunol 2001; 167: 2694-2699.
27 Andersen JF, Francischetti IM, Valenzuela JG. et al. Inhibition of hemostasis by a high affinity biogenic amine-binding protein from the saliva of a blood-feeding insect. J Biol Chem 2003; 278: 4611-4617.
28 Paesen GC, Adams PL, Harlos K. et al. Tick histamine-binding proteins: isolation, cloning, and three-dimensional structure. Mol Cell 1999; 03: 661-671.
29 Faili A, Randon J, Francischetti IM. et al. Convulxin-induced platelet aggregation is accompanied by a powerful activation of the phospholipase C pathway. Biochem J 1994; 298: 87-91.
30 Atkinson BT, Stafford MJ, Pears CJ. et al. Signalling events underlying platelet aggregation induced by the glycoprotein VI agonist convulxin. Eur J Biochem 2001; 268: 5242-5248.
31 Vargaftig BB, Chignard M, Benveniste J. Present concepts on the mechanisms of platelet aggregation. Biochem Pharmacol 1981; 30: 263-271.
32 Francischetti IM, Ghazaleh FA, Reis RA. et al. Convulxin induces platelet activation by a tyrosine-kinase-dependent pathway and stimulates tyrosine phosphorylation of platelet proteins, including PLC gamma 2, independently of integrin alpha IIb beta 3. Arch Biochem Biophys 1998; 353: 239-250.
33 Charo IF, Feinman RD, Detwiler TC. Interrelations of platelet aggregation and secretion. J Clin Invest 1977; 60: 866-873.
34 Mustard JF, Perry DW, Kinlough-Rathbone RL. et al. Factors responsible for ADP-induced release reaction of human platelets. Am J Physiol 1975; 228: 1757-1765.
35 Kunapuli SP. P2 receptors and platelet activation. SciWorld J 2002; 02: 424-433.
36 Packham MA, Bryant NL, Guccione MA. et al. Effect of the concentration of Ca2+ in the suspending medium on the responses of human and rabbit platelets to aggregating agents. Thromb Haemost 1989; 62: 968-976.
37 Jarvis GE, Atkinson BT, Snell DC. et al. Distinct roles of GPVI and integrin alpha(2)beta(1) in platelet shape change and aggregation induced by different collagens. Br J Pharmacol 2002; 137: 107-117.
38 Nakamura T, Jamieson GA, Okuma M. et al. Platelet adhesion to native type I collagen fibrils. Role of GPVI in divalent cation-dependent and -independent adhesion and thromboxane A2 generation. J Biol Chem 1998; 273: 4338-4344.
39 Huang JS, Ramamurthy SK, Lin X. et al. Cell signalling through thromboxane A2 receptors. Cell Signal 2004; 16: 521-533.
40 Morais Cabral JH, Atkins GL, Sanchez LM. et al. Arachidonic acid binds to apoli-poprotein D: implications for the protein's function. FEBS Lett 1995; 366: 53-56.
41 Ma D, Xu X, An S. et al. A novel family of RGD-containing disintegrins (Tablysin-15) from the salivary gland of the horsefly Tabanus yao targets alphaIIbbeta3 or alphaVbeta3 and inhibits platelet aggregation and angiogenesis. Thromb Haemost 2011; 105: 1032-1045.
42 Chmelar J, Oliveira CJ, Rezacova P. et al. A tick salivary protein targets cathepsin G and chymase and inhibits host inflammation and platelet aggregation. Blood 2011; 117: 736-744.
43 Li Z, Delaney MK, O'Brien KA. et al. Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol 2010; 30: 2341-2349.
44 Feletou M, Vanhoutte PM, Verbeuren TJ. The thromboxane/endoperoxide receptor (TP): the common villain. J Cardiovasc Pharmacol 2010; 55: 317-332.
45 Kahner BN, Shankar H, Murugappan S. et al. Nucleotide receptor signaling in platelets. J Thromb Haemost 2006; 04: 2317-2326.
46 Stafford MJ, Atkinson BT, Watson SP. et al. Signalling components underlying platelet aggregation to a Ca2+ ionophore and a phorbol ester. Platelets 2001; 12: 476-485.
47 Weech PK, Camato R, Milne RW. et al. Apolipoprotein D and cross-reacting human plasma apolipoproteins identified using monoclonal antibodies. J Biol Chem 1986; 261: 7941-7951.
48 Vogt M, Skerra A. Bacterially produced apolipoprotein D binds progesterone and arachidonic acid, but not bilirubin or E-3M2H. J Mol Recognit 2001; 14: 79-86.
49 Alvarenga PH, Francischetti IM, Calvo E. et al. The function and three-dimensional structure of a thromboxane A2/cysteinyl leukotriene-binding protein from the saliva of a mosquito vector of the malaria parasite. PLoS Biol 2010; 08: e1000547.
50 Ribeiro JM, Walker FA. High affinity histamine-binding and antihistaminic activity of the salivary nitric oxide-carrying heme protein (nitrophorin) of Rhodnius prolixus. J Exp Med 1994; 180: 2251-2257.