|
|
Prepublished online as a Blood First Edition Paper on April 30, 2002; DOI 10.1182/blood-2001-12-0339.
Submitted January 7, 2002
Accepted April 1, 2002
Sustained integrin ligation involves extracellular free sulfhydryls and enzymatically catalyzes disulfide exchange
Judith Lahav*, Kerstin Jurk, Oded Hess, Michael J Barnes, Richard W Farndale, Jacob Luboshitz, and Beate Kehrel
Hemostasis Laboratory, Rabin Medical Center-Beilinson Campus, Petah Tikva, Israel
Exp. und Klin Haemostaseology, Klinik und Poliklinik fuer Anaesthesiologie und operative Intensivmedizin, University of Muenster, Muenster, Germany
Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom
* Corresponding author; email: jlahav{at}netvision.net.il.
Recent observations suggest a pivotal role for free sulfhydryls in platelet integrin function and enzyme-mediated reduction of disulfide bonds on the platelet has been implicated. The platelet fibrinogen receptor  IIbß3 is the best-studied platelet integrin and serves as a model system for studying structure-function relationship of this family of adhesion receptors. The demonstration of free sulfhydryls on the exofacial domain of purified IIbß3, specifically in its activated conformation, prompted us to explore the potential for activation-dependent enzymatically-catalyzed thiol expression on the intact platelet and the possible role of surface associated protein disulfide isomerase (PDI) in IIbß3 ligation. Using the membrane-impermeant sulfhydryl blocker pCMPS, the inhibitor of disulfide exchange bacitracin and the monoclonal anti PDI antibody RL90, we probed fibrinogen binding to IIbß3 as well as ligation-induced allosteric changes in the conformation of IIbß3. We sought to distinguish possible involvement of disulfide exchange in agonist induced platelet stimulation from its role in integrin ligation. Analysis of the role of free thiols in platelet aggregation suggests a thiol-independent initial ligation followed by a thiol-dependent stabilization of binding. FACS analysis shows that sustained binding of fibrinogen as well as expression of LIBS epitopes and ligand-bound conformation depend on free thiols and on disulfide exchange. Expression of P-selectin is minimally affected even at complete inhibition of IIbß3 function.
These data indicate that while agonist-induced platelet stimulation is independent of ecto-sulfhydryls, engagement of integrin IIbß3 on the intact platelet is totally dependent on their enzymatically catalyzed surface expression.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
S. Carta, P. Castellani, L. Delfino, S. Tassi, R. Vene, and A. Rubartelli
DAMPs and inflammatory processes: the role of redox in the different outcomes
J. Leukoc. Biol.,
September 1, 2009;
86(3):
549 - 555.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Mor-Cohen, N. Rosenberg, M. Landau, J. Lahav, and U. Seligsohn
Specific Cysteines in {beta}3 Are Involved in Disulfide Bond Exchange-dependent and -independent Activation of {alpha}IIb{beta}3
J. Biol. Chem.,
July 11, 2008;
283(28):
19235 - 19244.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. R. Marwali, C.-P. Hu, B. Mohandas, A. Dandapat, P. Deonikar, J. Chen, I. Cawich, T. Sawamura, M. Kavdia, and J. L. Mehta
Modulation of ADP-Induced Platelet Activation by Aspirin and Pravastatin: Role of Lectin-Like Oxidized Low-Density Lipoprotein Receptor-1, Nitric Oxide, Oxidative Stress, and Inside-Out Integrin Signaling
J. Pharmacol. Exp. Ther.,
September 1, 2007;
322(3):
1324 - 1332.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. H. Versteeg and W. Ruf
Tissue Factor Coagulant Function Is Enhanced by Protein-disulfide Isomerase Independent of Oxidoreductase Activity
J. Biol. Chem.,
August 31, 2007;
282(35):
25416 - 25424.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. A. Jordan, J. M. Stevens, G. P. Hubbard, N. E. Barrett, T. Sage, K. S. Authi, and J. M. Gibbins
A role for the thiol isomerase protein ERP5 in platelet function
Blood,
February 15, 2005;
105(4):
1500 - 1507.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Krotz, H.-Y. Sohn, and U. Pohl
Reactive Oxygen Species: Players in the Platelet Game
Arterioscler Thromb Vasc Biol,
November 1, 2004;
24(11):
1988 - 1996.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. J. Forman, J. M. Fukuto, and M. Torres
Redox signaling: thiol chemistry defines which reactive oxygen and nitrogen species can act as second messengers
Am J Physiol Cell Physiol,
August 1, 2004;
287(2):
C246 - C256.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Chigaev, G. J. Zwartz, T. Buranda, B. S. Edwards, E. R. Prossnitz, and L. A. Sklar
Conformational Regulation of {alpha}4{beta}1-Integrin Affinity by Reducing Agents: "INSIDE-OUT" SIGNALING IS INDEPENDENT OF AND ADDITIVE TO REDUCTION-REGULATED INTEGRIN ACTIVATION
J. Biol. Chem.,
July 30, 2004;
279(31):
32435 - 32443.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Lahav, E. M. Wijnen, O. Hess, S. W. Hamaia, D. Griffiths, M. Makris, C. G. Knight, D. W. Essex, and R. W. Farndale
Enzymatically catalyzed disulfide exchange is required for platelet adhesion to collagen via integrin {alpha}2{beta}1
Blood,
September 15, 2003;
102(6):
2085 - 2092.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. A. Hotchkiss, A. W. Ashton, and E. L. Schwartz
Thymidine Phosphorylase and 2-Deoxyribose Stimulate Human Endothelial Cell Migration by Specific Activation of the Integrins {alpha}5{beta}1 and {alpha}V{beta}3
J. Biol. Chem.,
May 23, 2003;
278(21):
19272 - 19279.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
