Interaction of cryptogein with its binding sites in tobacco plasma

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Authors

SVOZILOVÁ Zuzana KAŠPAROVSKÝ Tomáš SKLÁDAL Petr LOCHMAN Jan

Year of publication 2009
Type Article in Periodical
Magazine / Source Analytical biochemistry
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords Membrane receptor Membrane vesicles Real time bioanalysis Elicitins Kinetic
Description Elicitins are low molecular weight proteins representing the elicitor family secreted by many species of the oomycete Phytophthora. Elicitins induce a hypersensitive reaction in tobacco, this process is triggered by binding of elicitin to the high affinity site on the plasma membrane. Specific interaction of cryptogein with the binding sites on tobacco plasma membranes was studied using the piezoelectric biosensor in real time in a flow-through mode. Cryptogeins (wild type and mutant forms) were covalently immobilized on the sensing surface and membrane vesicles s containing receptors were in solution. Kinetic characterization of the interaction provided values of kinetic rate association kas = 5.74x10E6 M1 s-1 and dissociation kdis = 6.87x10-4 s-1 constants, respectively. The kinetic equilibrium dissociation constant was calculated, KD = 12.0 nM. The piezoelectric biosensor appeared to be a convenient tool for studying interactions of receptors embedded in membrane vesicles.
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