PMID:2674127
| Citation |
Kuo, LC and Seaton, BA (1989) X-ray diffraction analysis on single crystals of recombinant Escherichia coli ornithine transcarbamoylase. J. Biol. Chem. 264:16246-8 |
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| Abstract |
Single crystals of recombinant Escherichia coli ornithine transcarbamoylase suitable for x-ray analysis have been grown from polyethylene glycol and 2-methyl-2,4-pentanediol. The space group has been determined as P3(1) or P3(2), with one protein trimer of three identical 36.8-kDa subunits in the asymmetric unit. The unit cell dimensions are a = b = 105.1 A and c = 87.8 A. The crystals diffract well to 3-A resolution and are quite resistant to radiation damage. Single crystals have also been grown of a genetically engineered site-specific mutant for which the replacement of an arginine (Arg-57) to a glycine has been shown to not only drastically affect the enzyme activity but also its kinetic mechanism (Kuo, L. C., Miller, A. W., Lee, S., and Kozuma, C. (1988) Biochemistry 27, 8823-8832). The crystals of the Arg-57----Gly mutant protein are isomorphous to those of the wild type. Crystal soaking experiments using both wild-type and Arg-57----Gly crystals in the presence of various ligands have provided evidence of specific conformational changes upon substrate binding which supports our previous kinetic and spectroscopic observations. |
| Links | |
| Keywords |
Crystallization; Escherichia coli/enzymology; Escherichia coli/genetics; Humans; Ligands; Ornithine Carbamoyltransferase/genetics; Protein Conformation; Recombinant Proteins; X-Ray Diffraction/methods |
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