title: Effect of flexibility and positive charge of the C-terminal domain on the activator P14K function for nitrile hydratase in Pseudomonas putida.
abstract: A self-subunit swapping chaperone is crucial for cobalt incorporation into nitrile hydratase. However, further information about its structural features is not available. The flexibility and positive charge of the C-terminal domain of the self-subunit swapping chaperone (P14K) of nitrile hydratase from Pseudomonas putida NRRL-18668 play an important role in cobalt incorporation. C-terminal domain truncation, alternation of C-terminal domain flexibility through mutant P14K(G86I), and elimination of the positive charge in the C-terminal domain sharply affected nitrile hydratase cobalt content and activity. The flexible, positively charged C-terminal domain most likely carries out an external action that allows a cobalt-free nitrile hydratase to overcome an energetic barrier, resulting in a cobalt-containing nitrile hydratase.
authors.affiliation: Key Laboratory of Industrial Biotechnology, School of Biotechnology, Jiangnan University, Wuxi, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Wenjing
authors.initials: W
authors.lastname: Cui
authors.forename: Zhongmei
authors.initials: Z
authors.lastname: Liu
authors.forename: Youtian
authors.initials: Y
authors.lastname: Cui
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Xia
authors.forename: Michihiko
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Zhemin
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003035:Cobalt / Q000378:metabolism; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000737:chemistry* / Q000378:metabolism*; D006836:Hydro-Lyases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D016958:Pseudomonas putida / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D020536:Enzyme Activators; D018832:Molecular Chaperones; D003035:Cobalt; D006836:Hydro-Lyases; C048831:nitrile hydratase
keywords: cobalt incorporation; enzyme activation; nitrile hydratase; protein expression