Bacitracin reveals a role for multiple thiol isomerases in platelet function

被引:20
作者
Robinson, A [1 ]
O'Neill, S [1 ]
Kiernan, AS [1 ]
O'Donoghue, N [1 ]
Moran, N [1 ]
机构
[1] Royal Coll Surgeons Ireland, Dept Clin Pharmacol, Dublin 2, Ireland
关键词
bacitracin; integrin; thiol isomerase; platelet;
D O I
10.1111/j.1365-2141.2005.05878.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The platelet-specific integrin alpha IIb beta 3 has endogenous thiol isomerase activity associated with the CXXC motifs within the beta subunit. Using a highly purified form of bacitracin, a thiol isomerase inhibitor, we now provide further evidence of the functional significance of this enzymatic activity in integrin activation. In addition, we demonstrate a role for multiple thiol isomerases in platelet function. This bacitracin prevented platelet aggregation to thrombin and collagen, and directly inhibited alpha IIb beta 3 activation, as detected by PAC-1 binding. In parallel, bacitracin inhibited the endogenous thiol isomerase activity of purified alpha IIb beta 3 with a 50% inhibitory concentration of 15(.)5 mu mol/l. In order to determine whether the effects of bacitracin are solely mediated by inhibition of integrin enzymatic activity, we examined integrin-independent indices of platelet activation. We found bacitracin inhibited both platelet secretion (CD62P and CD63) and thromboxane (TxA(2)) production, with complete inhibition at different concentrations. Thus, we demonstrated a role for multiple thiol isomerases in platelet function. Taken together, these studies support a role for the endogenous integrin thiol isomerase activity in activation of alpha IIb beta 3 and highlight the novel regulation of platelet function by other, as yet undefined thiol isomerases.
引用
收藏
页码:339 / 348
页数:10
相关论文
共 39 条
[21]   INHIBITION OF A REDUCTIVE FUNCTION OF THE PLASMA-MEMBRANE BY BACITRACIN AND ANTIBODIES AGAINST PROTEIN DISULFIDE-ISOMERASE [J].
MANDEL, R ;
RYSER, HJP ;
GHANI, F ;
WU, M ;
PEAK, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (09) :4112-4116
[22]   A palmitylated peptide derived from the glycoprotein lbβ cytoplasmic tail inhibits platelet activation [J].
Martin, K ;
Meade, G ;
Moran, N ;
Shields, DC ;
Kenny, D .
JOURNAL OF THROMBOSIS AND HAEMOSTASIS, 2003, 1 (12) :2643-2652
[23]   PURIFICATION OF NONANTIBIOTIC INSULINASE INHIBITORS FROM BACITRACIN [J].
MEDINA, V ;
KESNER, L ;
STRACHER, A .
BIOCHEMICAL MEDICINE AND METABOLIC BIOLOGY, 1993, 49 (02) :255-264
[24]   Metal binding and structure-activity relationship of the metalloantibiotic peptide bacitracin [J].
Ming, LJ ;
Epperson, JD .
JOURNAL OF INORGANIC BIOCHEMISTRY, 2002, 91 (01) :46-58
[25]   The platelet integrin αIIbβ3 has an endogenous thiol isomerase activity [J].
O'Neill, S ;
Robinson, A ;
Deering, A ;
Ryan, M ;
Fitzgerald, DJ ;
Moran, N .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (47) :36984-36990
[26]  
ONEILL S, 1999, THROMB DIATH HAEMO, V81, pA1561
[27]  
PEERSCHKE EI, 1995, THROMB HAEMOSTASIS, V73, P862
[28]  
PLOW EF, 1987, BLOOD, V70, P110
[29]   Enzyme destruction by a protease contaminant in bacitracin [J].
Rogelj, S ;
Reiter, KJ ;
Kesner, L ;
Li, M ;
Essex, D .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2000, 273 (03) :829-832
[30]   A point mutation in the cysteine-rich domain of glycoprotein (GP) IIIa results in the expression of a GPIIb-IIIa (αIIbβ3) integrin receptor locked in a high-affinity state and a Glanzmann thrombasthenia-like phenotype [J].
Ruiz, C ;
Liu, CY ;
Sun, QH ;
Sigaud-Fiks, M ;
Fressinaud, E ;
Muller, JY ;
Nurden, P ;
Nurden, AT ;
Newman, PJ ;
Valentin, N .
BLOOD, 2001, 98 (08) :2432-2441