An analysis of two refolding routes for a C-terminally truncated human collagenase-3 expressed in Escherichia coli.
We describe here the expression of a C-terminally truncated form of human procollagenase-3 in Escherichia coli. The protein was found almost exclusively in inclusion bodies that were solubilized and refolded by two separate methods and then purified on Ni-NTA agarose. The purified proenzyme could be activated with either trypsin or APMA and active enzyme could be purified on a peptidic hydroxamate affinity column. Competitive elution from the affinity matrix yielded a highly purified preparation.
Hardern
I M
IM
Enabling Science and Technology-Biology, AstraZeneca Pharmaceuticals, Mereside, Alderley Park, Macclesfield, Cheshire, SK10 4TG, United Kingdom.
Knauper
V
V
Ernill
R J
RJ
Taylor
I W
IW
Cooper
K L
KL
Abbott
W M
WM
D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D017364:Collagenases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D053509:Matrix Metalloproteinase 13; D017510:Protein Folding
D004792:Enzyme Precursors; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13