In the present study, surface plasmon resonance biosensor was employed to characterize the binding interaction between wild type porcine Pyridoxal Kinase (PK) and an immobilized substrate, pyridoxamine. The binding of PK to the immobilized pyridoxamine was followed in real time and the kinetic parameters were derived from the sensorgram using the BlAevaluationTM software. At pH 7.4, the association rate constant and dissociation rate constant of the wild-type PK for immobilized pyridoxamine are 7.25 x 103±0.35 x 103M-1s-1 and 5.01 x 10-3±0.93 x 10-3s-1respectively. The equilibrium affinity constant of the wild-type PK is 1.45 x 106M-1. The effect of buffer pH on the binding kinetic parameters was also determined.
|Title of host publication
|Biochemistry and molecular biology of vitamin B₆ and PQQ-dependent proteins
|Number of pages
|Published - 2000