[3Fe-4S] to [4Fe-4S] cluster conversion in Desulfovibrio fructosovorans [NiFe] hydrogenase by site-directed mutagenesis Export

@article{citeulike:4847997, abstract = {The role of the high potential [3Fe-4S] cluster of [NiFe] hydrogenase from species located halfway between the proximal and distal low potential [4Fe-4S] clusters has been investigated by using site-directed mutagenesis. Proline 238 of [NiFe] hydrogenase, which occupies the position of a potential ligand of the lacking fourth Fe-site of the [3Fe-4S] cluster, was replaced by a cysteine residue. The properties of the mutant enzyme were investigated in terms of enzymatic activity, EPR, and redox properties of the iron-sulfur centers and crystallographic structure. We have shown on the basis of both spectroscopic and x-ray crystallographic studies that the [3Fe-4S] cluster of hydrogenase was converted into a [4Fe-4S] center in the P238 mutant. The [3Fe-4S] to [4Fe-4S] cluster conversion resulted in a lowering of approximately 300 mV of the midpoint potential of the modified cluster, whereas no significant alteration of the spectroscopic and redox properties of the two native [4Fe-4S] clusters and the NiFe center occurred. The significant decrease of the midpoint potential of the intermediate Fe-S cluster had only a slight effect on the catalytic activity of the P238C mutant as compared with the wild-type enzyme. The implications of the results for the role of the high-potential [3Fe-4S] cluster in the intramolecular electron transfer pathway are discussed.}, author = {Rousset, Marc and Montet, Yael and Guigliarelli, Bruno and Forget, Nicole and Asso, Marcel and Bertrand, Patrick and Fontecilla-Camps, Juan C. and Hatchikian, E. Claude}, citeulike-article-id = {4847997}, citeulike-linkout-0 = {http://www.pnas.org/content/95/20/11625.long.abstract}, citeulike-linkout-1 = {http://www.pnas.org/content/95/20/11625.long.full.pdf}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/9751716}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=9751716}, day = {29}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {cluster, directed, fructosovorans, hydrogenase, iron, mutagenesis, nife, site, sulphur}, month = {September}, number = {20}, pages = {11625--11630}, posted-at = {2009-10-03 17:01:00}, priority = {2}, title = {[3Fe-4S] to [4Fe-4S] cluster conversion in Desulfovibrio fructosovorans [NiFe] hydrogenase by site-directed mutagenesis}, url = {http://www.pnas.org/content/95/20/11625.long.abstract}, volume = {95}, year = {1998} }

TY - JOUR ID - citeulike:4847997 L3 - citeulike-article-id:4847997 N2 - The role of the high potential [3Fe-4S] cluster of [NiFe] hydrogenase from species located halfway between the proximal and distal low potential [4Fe-4S] clusters has been investigated by using site-directed mutagenesis. Proline 238 of [NiFe] hydrogenase, which occupies the position of a potential ligand of the lacking fourth Fe-site of the [3Fe-4S] cluster, was replaced by a cysteine residue. The properties of the mutant enzyme were investigated in terms of enzymatic activity, EPR, and redox properties of the iron-sulfur centers and crystallographic structure. We have shown on the basis of both spectroscopic and x-ray crystallographic studies that the [3Fe-4S] cluster of hydrogenase was converted into a [4Fe-4S] center in the P238 mutant. The [3Fe-4S] to [4Fe-4S] cluster conversion resulted in a lowering of approximately 300 mV of the midpoint potential of the modified cluster, whereas no significant alteration of the spectroscopic and redox properties of the two native [4Fe-4S] clusters and the NiFe center occurred. The significant decrease of the midpoint potential of the intermediate Fe-S cluster had only a slight effect on the catalytic activity of the P238C mutant as compared with the wild-type enzyme. The implications of the results for the role of the high-potential [3Fe-4S] cluster in the intramolecular electron transfer pathway are discussed. IS - 20 JF - Proceedings of the National Academy of Sciences of the United States of America EP - 11630 TI - [3Fe-4S] to [4Fe-4S] cluster conversion in Desulfovibrio fructosovorans [NiFe] hydrogenase by site-directed mutagenesis VL - 95 SP - 11625 KW - cluster KW - directed KW - fructosovorans KW - hydrogenase KW - iron KW - mutagenesis KW - nife KW - site KW - sulphur AU - Rousset, Marc AU - Montet, Yael AU - Guigliarelli, Bruno AU - Forget, Nicole AU - Asso, Marcel AU - Bertrand, Patrick AU - Fontecilla-Camps, Juan AU - Hatchikian, Claude PY - 1998/09/29/ UR - http://www.pnas.org/content/95/20/11625.long.abstract ER -