Abstract
An Escherichia coli clone encoding a single cohesin domain of the cellulosome-integrating protein CipA from Clostridium thermocellum was constructed, and the corresponding polypeptide was purified, treated with papain, and crystallized from a PEG 8000 solution. Crystals exhibit orthorhombic symmetry, space group P2(1)2(1)2(1), with cell dimensions a = 37.7 A, b = 80.7 A, c = 93.3 A, and four or eight molecules in the unit cell. The crystals diffract X-rays to beyond 2 A resolution and are suitable for further crystallographic studies.
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- Fujino T., Béguin P., Aubert J. P. Cloning of a Clostridium thermocellum DNA fragment encoding polypeptides that bind the catalytic components of the cellulosome. FEMS Microbiol Lett. 1992 Jul 1;73(1-2):165–170. doi: 10.1016/0378-1097(92)90602-k. [DOI] [PubMed] [Google Scholar]
- Gerngross U. T., Romaniec M. P., Kobayashi T., Huskisson N. S., Demain A. L. Sequencing of a Clostridium thermocellum gene (cipA) encoding the cellulosomal SL-protein reveals an unusual degree of internal homology. Mol Microbiol. 1993 Apr;8(2):325–334. doi: 10.1111/j.1365-2958.1993.tb01576.x. [DOI] [PubMed] [Google Scholar]
- Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
- Salamitou S., Raynaud O., Lemaire M., Coughlan M., Béguin P., Aubert J. P. Recognition specificity of the duplicated segments present in Clostridium thermocellum endoglucanase CelD and in the cellulosome-integrating protein CipA. J Bacteriol. 1994 May;176(10):2822–2827. doi: 10.1128/jb.176.10.2822-2827.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tokatlidis K., Dhurjati P., Béguin P. Properties conferred on Clostridium thermocellum endoglucanase CelC by grafting the duplicated segment of endoglucanase CelD. Protein Eng. 1993 Nov;6(8):947–952. doi: 10.1093/protein/6.8.947. [DOI] [PubMed] [Google Scholar]
- Tokatlidis K., Salamitou S., Béguin P., Dhurjati P., Aubert J. P. Interaction of the duplicated segment carried by Clostridium thermocellum cellulases with cellulosome components. FEBS Lett. 1991 Oct 21;291(2):185–188. doi: 10.1016/0014-5793(91)81279-h. [DOI] [PubMed] [Google Scholar]