Analysis of Interaction between PHO4 and PHO2 Protein by Real Time BIA.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
; 31(2): 145-149, 1999.
Article
en En
| MEDLINE
| ID: mdl-12136204
ABSTRACT
Applying real time BIA(biomolecular interaction analysis) the interaction between yeast PHO4 and PHO2 protein was analyzed. Recombinant PHO4 protein was coupled at the sensor chip via amino group. 5 &mgr;mol/L of recombinant PHO2 and PHO2 mutants which were fused with glutathione S-transferase were injected respectively. The mutant whose Ser 230 was changed into Asp (GST-2SD) showed high SPR (surface plasmon resonance) signal while wild type PHO2 and the mutant where Ser 230 was changed into Ala (GST-2SA) did not. This result indicated that there was interaction between PHO4 and GST-2SD proteins and the phosphorylation of the Ser 230 of PHO2 was essential. Five different concentration of GST-2SD (0.5 0.75 1.0 1.5 2.0&mgr;mol/L) were injected separately and the interaction kinetics was analyzed. The association rate constant is 2.4x10(4)(mol/L)(-1)s(-1) dissociation rate constant is 3.5x10(-5)s(-1) and association constant is 6.9x10(8) (mol/L)(-1).
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Colección:
01-internacional
Base de datos:
MEDLINE
Idioma:
En
Revista:
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
Año:
1999
Tipo del documento:
Article