Iranian Journal of Biotechnology 2019-Sep
Role of Molecular Interactions and Oligomerization in Chaperone Activity of Recombinant Acr from Mycobacterium tuberculosis.
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Background
The chaperone activity of Mycobacterium tuberculosis Acr is an important function that helps to prevent misfolding of protein substrates inside the host, especially in conditions of hypoxia.Material and Methods
M. tuberculosis acr gene was cloned with an N-terminal His-tag in pET28a and expressed with IPTG induction in BL2 (DE3) competent Escherichia coli. The activity of a recombinant Acr without gel filtration was checked by preventing thermal aggregation of citrate synthase at 45°C and the chaperone activity against insulin B chain aggregation at 60°C and 37°C. On further purification using gel filtration chromatography, the protein was again tested for chaperone activity using insulin as substrate at 37°C with two types of samples without and with gel filtration designated A and B respectively. The effects of pre-heat treatment at 60 °C on chaperone activity of both A and B samples were studied by performing the chaperone assay at 37°C.