Determination of Kinetic Constants for the Interaction Between the Platelet Glycoprotein IIb-IIIa and Fibrinogen by Means of Surface Plasmon Resonance
| Autor: | Josef Hübscher, W. C. Kouns, Beat Steiner, Jurg Hurst, Daniel Schlatter, Richard Barner, Walter Huber |
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| Rok vydání: | 1995 |
| Předmět: |
Kinetics
Molecular Sequence Data Platelet Membrane Glycoproteins In Vitro Techniques Fibrinogen Platelet membrane glycoprotein Ligands Binding Competitive Biochemistry Dissociation (chemistry) medicine Humans Amino Acid Sequence Binding site Surface plasmon resonance chemistry.chemical_classification Chromatography Binding Sites Chemistry Dissociation constant Solubility Biophysics Thermodynamics Glycoprotein Oligopeptides medicine.drug |
| Zdroj: | European Journal of Biochemistry. 227:647-656 |
| ISSN: | 1432-1033 0014-2956 |
| Popis: | The binding reaction between purified human platelet glycoprotein IIb-IIIa and fibrinogen was investigated by real-time measurements using the surface-plasmon-resonance sensor technology. In these experiments, either glycoprotein IIb-IIIa or fibrinogen was immobilized on a sensor surface. The time-dependent change in surface coverage that occurred immediately upon contact with a solution of the complementary protein was then detected. The ability to record this dynamic event from its initiation allowed the collection of kinetic and thermodynamic data over an extended time period. These data indicated that initially, in fast reaction, a reversible low-affinity complex with an equilibrium dissociation constant, Kd, of 155-180 nM was formed. In a subsequent slower reaction this complex was transformed into a more stable high-affinity complex with a Kd of 20-70 nM. Efficient dissociation of the high-affinity complex could only be induced in the presence of a competitive inhibitor such as RGDV. These data demonstrate that the binding between glycoprotein IIb-IIIa and fibrinogen is not a single monophasic reaction, but is composed of at least two consecutive processes both with their own kinetics. |
| Databáze: | OpenAIRE |
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