Investigating intermolecular forces associated with thrombus initiation using optical tweezers

Maneesh Arya; Jose A. Lopez; Gabriel M. Romo; Jing-Fei Dong; Larry V. McIntire; Joel L. Moake; Bahman Anvari

doi:10.1117/12.468336

28 May 2002 Investigating intermolecular forces associated with thrombus initiation using optical tweezers

Maneesh Arya, Jose A. Lopez, Gabriel M. Romo, Jing-Fei Dong, Larry V. McIntire, Joel L. Moake, Bahman Anvari

Proceedings Volume 4622, Optical Diagnostics of Living Cells V; (2002) https://doi.org/10.1117/12.468336
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

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.

Citation Download Citation

Maneesh Arya, Jose A. Lopez, Gabriel M. Romo, Jing-Fei Dong, Larry V. McIntire, Joel L. Moake, and Bahman Anvari "Investigating intermolecular forces associated with thrombus initiation using optical tweezers", Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); https://doi.org/10.1117/12.468336

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