title: An analysis of two refolding routes for a C-terminally truncated human collagenase-3 expressed in Escherichia coli.
abstract: 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.
authors.affiliation: Enabling Science and Technology-Biology, AstraZeneca Pharmaceuticals, Mereside, Alderley Park, Macclesfield, Cheshire, SK10 4TG, United Kingdom.
authors.forename: I M
authors.initials: IM
authors.lastname: Hardern
authors.forename: V
authors.initials: V
authors.lastname: Knauper
authors.forename: R J
authors.initials: RJ
authors.lastname: Ernill
authors.forename: I W
authors.initials: IW
authors.lastname: Taylor
authors.forename: K L
authors.initials: KL
authors.lastname: Cooper
authors.forename: W M
authors.initials: WM
authors.lastname: Abbott
mesh_terms: 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
chemical_list: D004792:Enzyme Precursors; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13