Spore-coat laccase CotA from Bacillus subtilis: crystallization and preliminary X-ray characterization by the MAD method.
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.
Enguita
Francisco J
FJ
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2781-901 Oeiras, Portugal.
Matias
Pedro M
PM
Martins
Lígia O
LO
Plácido
Diana
D
Henriques
Adriano O
AO
Carrondo
Maria Arménia
MA
D001412:Bacillus subtilis / Q000201:enzymology* / Q000254:growth & development; D003460:Crystallization; D018360:Crystallography, X-Ray; D042845:Laccase; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D013171:Spores, Bacterial / Q000737:chemistry*
D010088:Oxidoreductases; D042845:Laccase