Thiosulfinates Inhibit Platelet Aggregation and Microparticle Shedding at a Calpain-dependent Step
Francine Rendu
1
U 428 INSERM, Faculté de Pharmacie, Université René Descartes Paris V, Paris
,
Brigitte Brohard-Bohn
1
U 428 INSERM, Faculté de Pharmacie, Université René Descartes Paris V, Paris
,
Sabine Pain
1
U 428 INSERM, Faculté de Pharmacie, Université René Descartes Paris V, Paris
,
Christilla Bachelot-Loza
1
U 428 INSERM, Faculté de Pharmacie, Université René Descartes Paris V, Paris
,
Jacques Auger
2
IRBI, Université François Rabelais, Tours, France
› InstitutsangabenWe are grateful to Dr. P. Marche for his critical reading of the manuscript. S.P. was a recipient of Ministère de l'Education Nationale et de la Recherche. The work was supported by a grant from ARC (FR: 9545).
Thiosulfinates (TSs) are sulfur compounds generated through the processing of different Allium species with antiplatelet property. To further define this platelet inhibitory effect we studied diallyl-TS (Al2TS), dipropyl-TS (Pr2TS), and dimethyl-TS (Me2TS) on platelet responses. The three TSs inhibited dose-dependent platelet aggregation, with IC50 values of 15 ± 2, 19 ± 2, and 9 ± 1 μM for Al2TS, Pr2TS and Me2TS, respectively. TSs had no effect on the expression of a platelet procoagulant surface, measured by flow cytometry as the binding of annexin V-FITC. They inhibited the microparticle shedding and clot retraction. Since the microparticle shedding is a calpain-activation dependent step, we assessed calpain activation by analysis of autoproteolysis in shorter active forms and by talin proteolysis in the presence of TSs. Calpain activation was inhibited by TSs independently of fibrinogen binding. Thus, TSs represent a new category of platelet inhibitors, acting on cal-pain-induced events.
Keywords
Allium -
calpain -
microparticles -
platelet aggregation -
talin
References
1
Wu KK.
Platelet activation mechanisms and markers in arterial thrombosis. J Int Med 1996; 239: 17-34.
5
Fox JEB,
Austin CD,
Boyles JK,
Steffen PK.
Role of the membrane skeleton in preventing the shedding of procoagulant-rich microvesicles from the platelet plasma membrane. J Cell Biol 1990; 111: 483-93.
10
Wencel-Drake JD,
Okita JR,
Annis DS,
Kunicki TJ.
Activation of calpain I and hydrolysis of calpain substrates (Actin-binding protein, glycoprotein Ib, and talin) are not a function of thrombin-induced platelet aggregation. Arterios Thromb 1991; 11: 882-91.
11
Auger J,
Lalau-Keraly FX,
Belinsky C.
Thiosulfinates in vapor phase are stable and they can persist in the environment of allium. Chemospher 1990; 21: 837-43.
13
Schoenwaelder SM,
Yuan Y,
Cooray P,
Salem HH,
Jackson SP.
Calpain cleavage of focal adhesion proteins regulates the cytoskeletal attachment of integrin αIIbβ3 (Platelet glycoprotein IIb/IIIa) and the cellular retraction of fibrin clots. J Biol Chem 1997; 272: 1694-702.
14
Dachary-Prigent J,
Freyssinet JM,
Pasquet JM,
Carron JC,
Nurden AT.
Annexin V as a probe of aminophospholipid exposure and platelet membrane vesiculation: a flow cytometry study showing a role for free sulfhydryl groups. Blood 1993; 81: 2554-65.
15
Dachary-Prigent J,
Pasquet JM,
Freyssinet JM,
Nurden AT.
Calcium involvement in aminophospholipid exposure and microparticle formation during platelet activation: a study using Ca2+-ATPase inhibitors. Biochemistry 1995; 34: 11625-34.
16
Inomata M,
Hayashi M,
Ohno-Iwashita Y,
Tsubuki S,
Saido TC,
Kawashima S.
Involvement of calpain in integrin-mediated signal transduction. Arch Biochem Biophys 1996; 328: 129-34.
17
Fox JEB,
Taylor RG,
Taffarel M,
Boyles JK,
Goll DE.
Evidence that activation of platelet calpain is induced as a consequence of binding of adhesive ligand to the integrin, glycoprotein IIb-IIIa. J Cell Biol 1993; 120: 1501-7.
19
Apitz-Castro R,
Jain MK,
Bartoli F,
Ledezma E,
Ruiz MC,
Salas R.
Evidence for direct coupling of primary agonist-receptor interaction to the exposure of functional IIb-IIIa complexes in human blood platelets. Results from studies with the antiplatelet compound ajoene. Biochim Biophys Acta 1991; 1094: 269-80.
20
Apitz-Castro R,
Ledezma E,
Escalante J,
Jain MK.
The molecular basis of the antiplatelet action of ajoene: direct interaction with fibrinogen. Biochem Biophys Res Commun 1987; 144: 1348-50.
22
Srivastava KC,
Tyagi OD.
Effects of a garlic-derived principle (ajoene) on aggregation and arachidonic acid metabolism in human blood platelets. Prost Leuk Ess Fatty Acids 1993; 49: 587-95.
23
Apitz-Castro R,
Ledezma E,
Escalante J,
Jorquera A,
Pinate FM,
MorenoRea J,
Carillo G,
Leal O,
Jain MK.
Reversible prevention of platelet activation by (E,Z)-4,5,9-Trithiadodeca-1,6,11-triene 9-oxide (ajoene) in dogs under extracoporeal circulation. Drug Res 1988; 38: 901-4.
24
Bassé F,
Gaffet P,
Bienvenüe A.
Correlation between inhibition of cyto-skeleton proteolysis and anti-vesiculation effect of calpeptin during A23187-induced activation of human platelets: are vesicles shed by filopod fragmentation?. Biochim Biophys Acta 1994; 1190: 217-24.
25
Frangioni JV,
Oda A,
Smith M,
Salzman EW,
Neel BG.
Calpain-catalyzed cleavage and subcellular relocation of protein phosphotyrosine phosphatase 1B (PTP-1B) in human platelets. Embo J 1993; 12: 4843-56.
26
Oda A,
Druker BJ,
Ariyoshi H,
Smith M,
Salzman EW.
pp60src is an endogenous substrate for calpain in human blood platelets. J Biol Chem 1993; 268: 12603-8.
27
Schoenwaelder SM,
Kulkarni S,
Salem HH,
Imajoh-Ohmi S,
Yamao-Harigaya W,
Saido TC,
Jackson SP.
Distinct substrate specificities and functional roles for the 78- and 76-kDa forms of μ-calpain in human platelets. J Biol Chem 1997; 272: 24876-84.
29
Nomura S,
Komiyama Y,
Matsuura E,
Lan Xie G,
Katsura K,
Miyake T,
Miyasaki Y,
Kagawa H,
Koike T,
Fukuhara S.
Participation of αIIbβ3 in platelet microparticle generation by collagen plus thrombin. Haemostasis 1996; 26: 31-7.
32
Hayashi M,
Suzuki H,
Kawashima S,
Saido TC,
Inomata M.
The behavior of calpain-generated N- and C-terminal fragments of talin in integrin-mediated signaling pathways. Arch Biochem Biophys 1999; 371: 133-41.
33
Mondoro TH,
White MM,
Jennings LK.
Active GPIIb-IIIa conformations that link ligand interaction with cytoskeletal reorganization. Blood 2000; 96: 2487-95.
34
Cohen I,
Burk DL,
and White JG.
The effect of peptides and monoclonal antibodies that bind to platelet glycoprotein IIb-IIIa complex on the development of clot tension. Blood 1989; 73: 1880-7.
35
Wolf BB,
Goldstein JC,
Stennicke HR,
Beere H,
Amarante-Mendes GP,
Salvesen GS,
Green DR.
Calpain functions in a caspase-independent manner to promote apoptosis-like events during platelet activation. Blood 1999; 94: 1683-92.
37
Rabinkov A,
Miron T,
Konstantinovski L,
Wilchek M,
Mirelman D,
Weiner L.
The mode of action of allicin: trapping of radicals and interaction with thiol containing proteins. Biochim Biophys Acta 1998; 1379: 233-44.
38
Teyssier C,
Guenot L,
Suschetet M,
Siess MH.
Metabolism of diallyl disulfide by human liver microsomal cytochromes P-450 and flavin-containing monooxygenases. Drug Metab Dispos 1998; 27: 835-41.
40
Johnson MG,
Vaughn RH.
Death of Salmonella typhymurium and Escherichia coli in the presence of freshly reconstituted garlic and onion. Appl Microbiol 1969; 17: 903-5.
41
Ferary S,
Auger J.
What is the true odour of cut Allium? Complementarity of various hyphenated methods: gas chromatography-mass spectrometry and high-performance liquid chromatography-mass spectrometry with particle beam and atmospheric pressure ionization interfaces in sulphenic acids rearrangement components discrimination. J Chromat 1996; 750: 63-74.
42
Ferary S,
Keller J,
Boscher J,
Auger J.
Fast narrow-bore HPLC-DAD analysis of biologically active thiosulfinates obtained without solvent from some wild Allium species. Biomed Chromatogr 1998; 12: 104-6.
