PMID:1960717
| Citation |
McGrath, ME, Erpel, T, Browner, MF and Fletterick, RJ (1991) Expression of the protease inhibitor ecotin and its co-crystallization with trypsin. J. Mol. Biol. 222:139-42 |
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| Abstract |
We have expressed the serine protease inhibitor ecotin to high levels (greater than 400 mg/l of cell culture) in its natural mileau, the Escherichia coli periplasm, using the endogenous signal peptide and the heterologous tac promoter. After induction, functional, soluble ecotin comprises 15% of total cellular protein. This expression system has facilitated initiation of a crystallographic study to determine the structural basis for inhibition of the pancreatic serine proteases by ecotin. Ecotin was co-crystallized with rat trypsin mutant D102N. Preliminary crystallographic analysis of co-crystals showed that they diffract to at least 2.7 A, and indicate that they belong to the monoclinic space group, P21. The cell constants are a = 52.0 A, b = 93.3 A, c = 160.7 A, and beta = 96 degrees. Four molecules each of trypsin and ecotin are found in the asymmetric unit. |
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| Keywords |
Bacterial Proteins/chemistry; Bacterial Proteins/ultrastructure; Crystallography; Escherichia coli/enzymology; Escherichia coli Proteins; Periplasmic Proteins; Protease Inhibitors/chemistry; Protein Conformation; Recombinant Proteins/chemistry; Recombinant Proteins/ultrastructure; Trypsin/chemistry; Trypsin/ultrastructure |
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