Mutant human protein disulfide isomerase assists protein folding in a chaperone-like fashion.
Human protein disulfide isomerase with an extra 10 amino acid residues of AEITRIDPAM at the N-terminal was expressed in E. coli as a soluble protein comprising 20% of total cell proteins, and was purified to near homogeneity through one step of DEAE-Sephacel chromatography. The mutant enzyme, which had the same CD spectrum and comparable disulfide isomerase and thiol-protein oxidoreductase activities with that of the wild type human and bovine protein disulfide isomerases, also showed chaperone-like activity in stimulating the refolding of proteins containing no disulfide bond. The overall yield of the active product is about 20 mg 1-1 culture.
Gao
Y
Y
National Laboratory of Biomacromolecules, Academia Sinica, Beijing, People's Republic of China.
Quan
H
H
Jiang
M
M
Dai
Y
Y
Wang
C C
CC
D000595:Amino Acid Sequence; D001483:Base Sequence; D018833:Chaperonins / Q000378:metabolism*; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006801:Humans; D007535:Isomerases / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D019704:Protein Disulfide-Isomerases; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D018833:Chaperonins; D007535:Isomerases; D019704:Protein Disulfide-Isomerases