tailieunhanh - Báo cáo khoa học: Comparing the substrate specificities of cytochrome c biogenesis Systems I and II

c-Type cytochromes require specific post-translational protein systems, which vary in different organisms, for the characteristic covalent attach-ment of heme to the cytochrome polypeptide. Cytochromec biogenesis System II, found in chloroplasts and many bacteria, comprises four subun-its, two of which (ResB and ResC) are the minimal functional unit. | Comparing the substrate specificities of cytochrome c biogenesis Systems I and II Bioenergetics Alan D. Goddard Julie M. Stevens Arnaud Rondelet Elena Nomerotskaia James W. A. Allen and Stuart J. Ferguson Department of Biochemistry University of Oxford UK Keywords cytochrome c cytochrome c maturation heme heme provision System II Correspondence S. J. Ferguson Department of Biochemistry University of Oxford South Parks Road Oxford OX1 3QU UK Fax 44 1865 613201 Tel 44 1865 613299 E-mail These authors contributed equally to this work Received 28 July 2009 revised 12 October 2009 accepted 25 November 2009 doi c-Type cytochromes require specific post-translational protein systems which vary in different organisms for the characteristic covalent attachment of heme to the cytochrome polypeptide. Cytochrome c biogenesis System II found in chloroplasts and many bacteria comprises four subunits two of which ResB and ResC are the minimal functional unit. The ycf5 gene from Helicobacter pylori encodes a fusion of ResB and ResC. Heterologous expression of ResBC in Escherichia coli lacking its own biogenesis machinery allowed us to investigate the substrate specificity of System II. ResBC is able to attach heme to monoheme c-type cytochromes c550 from Paracoccus denitrificans and c552 from Hydrogenobacter thermophilus both normally matured by System I. The production of holocyto-chrome is enhanced by the addition of exogenous reductant. Single-cysteine variants of these cytochromes were not efficiently matured by System II but System I was able to produce detectable amounts of AXXCH variants this adds to evidence that there is no obligate requirement for a disulfide-bonded intermediate for the latter c-type cytochrome biogenesis system. In addition System II was able to mature an AXXAH-containing variant into a b-type cytochrome with implications for both heme supply to the periplasm and substrate recognition by .

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