An Efficient System for the Expression and Purification of Yeast Geranylgeranyl Protein Transferase Type I
To purify the geranylgeranyl protein transferase type I (GGPT-I) efficiently, a gene expression system using the pGEX-4T-1 vector was constructed. The cal 1 gene, encoding the β subunit of GGPT-I, was subcloned into the pGEX-4T-1 vector and co-transformed into E. coli cells harboring the ram2 gene,...
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Published in | BMB reports Vol. 31; no. 1; pp. 77 - 82 |
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Main Authors | , , |
Format | Journal Article |
Language | Korean |
Published |
생화학분자생물학회
30.01.1998
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Online Access | Get full text |
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Summary: | To purify the geranylgeranyl protein transferase type I (GGPT-I) efficiently, a gene expression system using the pGEX-4T-1 vector was constructed. The cal 1 gene, encoding the β subunit of GGPT-I, was subcloned into the pGEX-4T-1 vector and co-transformed into E. coli cells harboring the ram2 gene, the α subunit gene of GGPT-I. GGPT-I was highly expressed as a fusion protein with glutathione S-transferase (GST) in E. coli, purified to homogeneity by glutathione-agarose affinity chromatography, and the GST moiety was excised by thrombin treatment. The purified yeast GGPT-I showed a dose-dependent increase in the transferase activity, and its apparent K_m value for an undecapeptide fused with GST (GST-PEP) was 0.66μM and the apparent K_m value for geranylgeranyl pyrophosphate (GGPP) was 0.071 μM. |
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Bibliography: | Korean Society for Biochemistry and Molecular Biology |
ISSN: | 1976-6696 |