title: Effect of alteration of the C-terminal extension on the maturation and folding of the large subunit of the Escherichia coli hydrogenase-2.
abstract: Large subunits of NiFe-hydrogenases undergo a unique maturation process in which the last step consists of the endoproteolytic cleavage of the C-terminal extension after the Ni-Fe metal center has been assembled. To assess in vivo the influence of alteration of the C-terminal extension on the processing, green fluorescence protein (GFP) was fused to the C-terminus of the large subunit (HybC) of the Escherichia coli hydrogenase 2. Interestingly, no processing of HybC-GFP was observed. In addition, the chromophore of GFP was not formed, implying a nonproductive folding of the upstream HybC moiety. These results strongly suggest that the alteration of the C-terminus of the hydrogenase 2 large subunit interferes with the folding and processing of HybC.
authors.affiliation: Laboratoire de Chimie Bactérienne, UPR9043 CNRS, Institut de Biologie Structurale et Microbiologie, 31 chemin Joseph Aiguier, 13402 Marseille cedex 20, France.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.forename: Nathalie
authors.initials: N
authors.lastname: Pradel
authors.forename: Marie-Andrée
authors.initials: MA
authors.lastname: Mandrand-Berthelot
authors.forename: Zengliang
authors.initials: Z
authors.lastname: Yu
authors.forename: Long-Fei
authors.initials: LF
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006864:Hydrogenase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009532:Nickel / Q000378:metabolism; D017510:Protein Folding; D021122:Protein Subunits / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D009532:Nickel; D006864:Hydrogenase; C481721:hydrogenase 2, E coli