Acta Crystallographica Section D

Biological Crystallography

Volume 58, Part 9 (September 2002)

crystallization papers

Acta Cryst. (2002). D58, 1490-1493 [ doi:10.1107/S0907444902011575 ]

Spore-coat laccase CotA from Bacillus subtilis: crystallization and preliminary X-ray characterization by the MAD method

F. J. Enguita, P. M. Matias, L. O. Martins, D. Plácido, A. O. Henriques and M. A. Carrondo

Abstract: Bacterial endospores are highly resistant structures that allow survival for long periods of time in adverse environments. The spore-forming Gram-positive bacterium Bacillus subtilis synthesizes a coat around the endospore during development composed of several assembled polypeptides. The role of these components of the spore coat remains unclear; however, some of them appear to be enzymes possibly involved in the assembly process or in the final properties of the spore. The outer spore-coat protein CotA is a 65 kDa polypeptide showing a high degree of sequence similarity with copper-dependent oxidases, including fungal and plant laccases, ascorbate oxidase and CueO from Esherichia coli. CotA has been recently characterized as a copper-dependent laccase. Unlike previously reported laccases, CotA shows increased thermostability. Here, the crystallization of a recombinant CotA protein produced in E. coli and the preliminary characterization of the crystals is reported. Structure solution by the MAD method at the copper K edge is also reported.

Keywords: spore-coat laccase.