Purification and C-terminal domain comparison of heat shock protein 90(HSP90) in Candida albicans and human

Abstract

Abstract:

Objective To express and purify Candida albicans HSP90 (CaHSP90) and human HSP90 (hHSP90) and to compare the different effects of C-terminal domain on their oligomeric forms in vitro.Method The encoding sequences for wild type CaHSP90,hHSP90 and their C-terminal enzyme digested base sequences were amplified via PCR and the products were cloned into pET-22b⁺ vector using NdeI/XhoI cloning sites.The plasmids were expressed in E. coli BL21 (DE3) cells and the His-tagged proteins were purified through Ni²⁺-NTA column and Superdex-200 column chromatography on FPLC. The molecular weights and oligomeric forms of these proteins were analyzed by SDS-PAGE and size exclusion chromatography (SEC).Results All plasmids were successfully constructed by DNA sequencing and the proteins were pure enough for SEC analysis.The results showed that wild type CaHSP90 and hHSP90 have similar oligomeric forms,while their C-terminal mutants were different.Conclusion The C-terminal domain of CaHSP90 and hHSP90 have different effect on their oligomeric forms in vitro and might be a potential target for CaHSP90 inhibitor.

Key words: Candida albicans, HSP90, protein structure, protein expression and purification

CLC Number:

R379.4

MENG Jing-shun, WU Jian-hua, ZHAO Jing, WANG Yu-lian, XUE Wei. Purification and C-terminal domain comparison of heat shock protein 90(HSP90) in Candida albicans and human[J]. Chinese Journal of Mycology, 2016, 11(6): 321-326.

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