Kinetics of the interaction of desAABB-fibrin monomer with immobilized fibrinogen.
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).
Chtcheglova
Lilia A
LA
Living Matter Physics Laboratory, Institute of Physics of the Complex Matter, Swiss Federal Institute of Technology Lausanne (EPFL), CH-1015 Lausanne, Switzerland. lilia.chtcheglova@jku.at
Vogel
Monique
M
Gruber
Hermann J
HJ
Dietler
Giovanni
G
Haeberli
André
A
D001704:Biopolymers / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques; D005337:Fibrin / Q000737:chemistry / Q000378:metabolism*; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
D001704:Biopolymers; D010446:Peptide Fragments; D005337:Fibrin; D005340:Fibrinogen; C065735:glycyl-prolyl-arginyl-valyl-valyl-glutamic acid