TAILIEUCHUNG - Báo cáo khoa học: In vivo production of catalase containing haem analogues

Haem (protohaem IX) analogues are toxic compounds and have been con-sidered for use as antibacterial agents, but the primary mechanism behind their toxicity has not been demonstrated. Using the haem protein catalase in the Gram-positive bacterium Enterococcus faecalisas an experimental system, we show that a variety of haem analogues can be taken up by bac-terial cells and incorporated into haem-dependent enzymes. | ỊFEBS Journal In vivo production of catalase containing haem analogues Myriam Brugna Lena Tassef and Lars Hederstedt Microbiology Group Department of Biology Lund University Sweden Keywords antibacterial agents catalase Enterococcus faecalis haem protein metalporphyrins Correspondence L. Hederstedt Department of Biology Biology Building Lund University Solvegatan 35 SE-22362 Lund Sweden Fax 46 46 222 41 13 Tel 46 46 222 86 22 E-mail Present address Laboratoire de Bioénergétique et Ingenierie des Proteines Institut de la Mediterranee CNRS Marseille cedex 20 France Universite de Provence 3 Place Victor Hugo Marseille cedex 3 France Laboratoire des Interactions Plantes-Micro-organismes UMR INRA-CNRS Chemin de Borde-Rouge BP 52627 Castanet-Tolosan France Received 4 March 2010 revised 31 March 2010 accepted 9 April2010 Haem protohaem IX analogues are toxic compounds and have been considered for use as antibacterial agents but the primary mechanism behind their toxicity has not been demonstrated. Using the haem protein catalase in the Gram-positive bacterium Enterococcus faecalis as an experimental system we show that a variety of haem analogues can be taken up by bacterial cells and incorporated into haem-dependent enzymes. The resulting cofactor-substituted proteins are dysfunctional generally resulting in arrested cell growth or death. This largely explains the cell toxicity of haem analogues. In contrast to many other organisms E. faecalis does not depend on haem for growth and therefore resists the toxicity of many haem analogues. We have exploited this feature to establish a bacterial in vivo system for the production of cofactor-substituted haem protein variants. As a pilot study we produced isolated and analysed novel catalase variants in which the iron atom of the haem prosthetic group is replaced by other metals . cobalt gallium tin and zinc and also variants containing meso-protoheme IX ruthenium meso-protoporphyrin IX and .

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