Expression and Functional Analysis of Yeast PHO81 Ankyrin Repeats.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
; 31(2): 185-190, 1999.
Article
en En
| MEDLINE
| ID: mdl-12136213
ABSTRACT
There are six ankyrin repeats in yeast transcriptional factor PHO81. The PHO81 ankyrin repeats fused with glutathione S-transferase (GST-ANK) was overexpressed in E. coli and it existed in inclusion body form. Using reduced and oxidized glutathione systems the fused protein fragment was denatured and renatured and the soluble protein was obtained. Then it was purified by affinity purification through glutathione sepharose column and its activity was analyzed. The PHO85-PHO80 and PHO85-PAP1 kinase complexes were prepared via coimmunoprecipitation respectively. With purified recombinant PHO4 protein as substrate of the kinase complexes when the purified GST-ANK was added the kinase activity of PHO85-PHO80 or PHO85-PAP1 kinase complex was inhibited. These studies indicate that the ankyrin repeats interact with PHO85-PHO80 or PHO85-PAP1 complex inhibit their kinase activities and are crucial in interaction between PHO81 and other proteins.
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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