The plasma protein β2GPI (β2-glycoprotein I) has been proposed to mediate phagocytosis of apoptotic cells and to play a role in the antiphospholipid syndrome. This suggestion is based mainly on the presumption that β2GPI has an appreciable interaction with PS (phosphatidylserine)-exposing cell membranes. However, quantitative data on the binding of β2GPI to PS-exposing cells under physiologically relevant conditions are scarce and conflicting. Therefore we evaluated the binding of β2GPI to PS-expressing blood platelets. Flow cytometry showed that binding of β2GPI is negligible at physiological ionic strength, in contrast with significant binding occurring at low ionic strength. Binding parameters of β2GPI and (for comparison) prothrombin were quantified by ellipsometric measurement of protein depletion from the supernatant following incubation with platelets. At low ionic strength (20 mM NaCl, no CaCl2), a dissociation constant (Kd) of 0.2 μM was found for β2GPI, with 7.4×105 binding sites per platelet. Under physiologically relevant conditions (120 mM NaCl and 3 mM CaCl2), binding of β2GPI was not detectable (extrapolated Kd>80 μM). Prothrombin binding (at 3 mM CaCl2) was much less affected by ionic strength: Kd values of 0.5 and 1.4 μM were observed at 20 and 120 mM NaCl respectively. The low affinity and the presence of many lipid-binding proteins in plasma that can compete with the binding of β2GPI suggest that only a small fraction (<5%) of the binding sites on PS-exposing blood cells are likely to be occupied by β2GPI. These findings are discussed in relation to the alleged (patho-)physiological functions of β2GPI.
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Research Article|
February 22 2005
Quantitative determination of the binding of β2-glycoprotein I and prothrombin to phosphatidylserine-exposing blood platelets
Edouard M. BEVERS;
Edouard M. BEVERS
1
*Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
1To whom correspondence should be addressed (email em.bevers@bioch.unimaas.nl).
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Marie P. JANSSEN;
Marie P. JANSSEN
*Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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Paul COMFURIUS;
Paul COMFURIUS
*Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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Krishnakumar BALASUBRAMANIAN;
Krishnakumar BALASUBRAMANIAN
†Department of Cancer Biology, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, U.S.A.
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Alan J. SCHROIT;
Alan J. SCHROIT
†Department of Cancer Biology, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, U.S.A.
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Robert F. A. ZWAAL;
Robert F. A. ZWAAL
*Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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George M. WILLEMS
George M. WILLEMS
*Cardiovascular Research Institute Maastricht, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands
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Publisher: Portland Press Ltd
Received:
July 09 2004
Revision Received:
September 17 2004
Accepted:
November 02 2004
Accepted Manuscript online:
November 02 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 386 (2): 271–279.
Article history
Received:
July 09 2004
Revision Received:
September 17 2004
Accepted:
November 02 2004
Accepted Manuscript online:
November 02 2004
Citation
Edouard M. BEVERS, Marie P. JANSSEN, Paul COMFURIUS, Krishnakumar BALASUBRAMANIAN, Alan J. SCHROIT, Robert F. A. ZWAAL, George M. WILLEMS; Quantitative determination of the binding of β2-glycoprotein I and prothrombin to phosphatidylserine-exposing blood platelets. Biochem J 1 March 2005; 386 (2): 271–279. doi: https://doi.org/10.1042/BJ20041167
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