Hardern I M, Knauper V, Ernill R J, Taylor I W, Cooper K L, Abbott W M
Enabling Science and Technology-Biology, AstraZeneca Pharmaceuticals, Mereside, Alderley Park, Macclesfield, Cheshire, SK10 4TG, United Kingdom.
Protein Expr Purif. 2000 Jul;19(2):246-52. doi: 10.1006/prep.2000.1244.
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.
我们在此描述了人原胶原酶-3 C 末端截短形式在大肠杆菌中的表达。该蛋白几乎完全存在于包涵体中，通过两种不同方法将其溶解并复性，然后在镍-亚氨基二乙酸琼脂糖上进行纯化。纯化的酶原可用胰蛋白酶或氨基苯汞乙酸激活，活性酶可在肽基羟肟酸亲和柱上进行纯化。从亲和基质上竞争性洗脱得到了高度纯化的制剂。
