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Proceedings Paper

Investigating intermolecular forces associated with thrombus initiation using optical tweezers
Author(s): Maneesh Arya; Jose A. Lopez; Gabriel M. Romo; Jing-Fei Dong; Larry V. McIntire; Joel L. Moake; Bahman Anvari
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Paper Abstract

Thrombus formation occurs when a platelet membrane receptor, glycoprotein (GP) Ib-IX-V complex, binds to its ligand, von Willebrand factor (vWf), in the subendothelium or plasma. To determine which GP Ib-IX-V amino acid sequences are critical for bond formation, we have used optical tweezers to measure forces involved in the binding of vWf to GP Ib-IX-V variants. Inasmuch as GP Ib(alpha) subunit is the primary component in human GP Ib-IX-V complex that binds to vWf, and that canine GP Ib(alpha) , on the other hand, does not bind to human vWf, we progressively replaced human GP Ib(alpha) amino acid sequences with canine GP Ib(alpha) sequences to determine the sequences essential for vWf/GP Ib(alpha) binding. After measuring the adhesive forces between optically trapped, vWf-coated beads and GP Ib(alpha) variants expressed on mammalian cells, we determined that leucine- rich repeat 2 of GP Ib(alpha) was necessary for vWf/GP Ib-IX- V bond formation. We also found that deletion of the N- terminal flanking sequence and leucine-rich repeat 1 reduced adhesion strength to vWf but did not abolish binding. While divalent cations are known to influence binding of vWf, addition of 1mM CaCl2 had no effect on measured vWf/GP Ib(alpha) bond strengths.

Paper Details

Date Published: 28 May 2002
PDF: 11 pages
Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); doi: 10.1117/12.468336
Show Author Affiliations
Maneesh Arya, Rice Univ. (United States)
Jose A. Lopez, Baylor College of Medicine (United States)
Gabriel M. Romo, Baylor College of Medicine (United States)
Jing-Fei Dong, Baylor College of Medicine (United States)
Larry V. McIntire, Rice Univ. (United States)
Joel L. Moake, Rice Univ. and Baylor College of Medicine (United States)
Bahman Anvari, Rice Univ. (United States)

Published in SPIE Proceedings Vol. 4622:
Optical Diagnostics of Living Cells V
Robert C. Leif; Daniel L. Farkas; Robert C. Leif, Editor(s)

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