desAABB - 纤维蛋白单体与固定化纤维蛋白原相互作用的动力学

Kinetics of the interaction of desAABB-fibrin monomer with immobilized fibrinogen.

作者信息

Chtcheglova Lilia A, Vogel Monique, Gruber Hermann J, Dietler Giovanni, Haeberli André

机构信息

Living Matter Physics Laboratory, Institute of Physics of the Complex Matter, Swiss Federal Institute of Technology Lausanne (EPFL), CH-1015 Lausanne, Switzerland.

出版信息

Biopolymers. 2006 Sep;83(1):69-82. doi: 10.1002/bip.20529.

DOI:10.1002/bip.20529

PMID:16639743

Abstract

The soluble and stable fibrin monomer-fibrinogen complex (SF) is well known to be present in the circulating blood of healthy individuals and of patients with thrombotic diseases. However, its physiological role is not yet fully understood. To deepen our knowledge about this complex, a method for the quantitative analysis of interaction between soluble fibrin monomers and surface-immobilized fibrinogen has been established by means of resonant mirror (IAsys) and surface plasmon resonance (BIAcore) biosensors. The protocols have been optimized and validated by choosing appropriate immobilization procedures with regeneration steps and suitable fibrin concentrations. The highly specific binding of fibrin monomers to immobilized fibrin(ogen), or vice versa, was characterized by an affinity constant of approximately 10(-8)M, which accords better with the direct dissociation of fibrin triads (KD approximately 10(-8) -10(-9) M) (J. R. Shainoff and B. N. Dardik, Annals of the New York Academy of Science, 1983, Vol. 27, pp. 254-268) than with earlier estimations of the KD for the fibrin-fibrinogen complex (KD approximately 10(-6) M) (J. L. Usero, C. Izquierdo, F. J. Burguillo, M. G. Roig, A. del Arco, and M. A. Herraez, International Journal of Biochemistry, 1981, Vol. 13, pp. 1191-1196).

摘要

可溶性稳定纤维蛋白单体 - 纤维蛋白原复合物（SF）在健康个体和血栓性疾病患者的循环血液中均有存在，这是众所周知的。然而，其生理作用尚未完全明确。为了深入了解这种复合物，借助共振镜（IAsys）和表面等离子体共振（BIAcore）生物传感器建立了一种定量分析可溶性纤维蛋白单体与表面固定的纤维蛋白原之间相互作用的方法。通过选择带有再生步骤的合适固定程序和合适的纤维蛋白浓度，对实验方案进行了优化和验证。纤维蛋白单体与固定化纤维蛋白（原）的高度特异性结合，其特征在于亲和常数约为10^(-8)M，这与纤维蛋白三联体的直接解离（KD约为10^(-8) - 10^(-9) M）（J.R. Shainoff和B.N. Dardik，《纽约科学院学报》，1983年，第27卷，第254 - 268页）更为相符，而与早期对纤维蛋白 - 纤维蛋白原复合物KD的估计值（KD约为10^(-6) M）（J.L. Usero、C. Izquierdo、F.J. Burguillo、M.G. Roig、A. del Arco和M.A. Herraez，《国际生物化学杂志》，1981年，第13卷，第1191 - 1196页）不同。