Effect of flexibility and positive charge of the C-terminal domain on the activator P14K function for nitrile hydratase in Pseudomonas putida

FEMS Microbiol Lett. 2014 Mar;352(1):38-44. doi: 10.1111/1574-6968.12376. Epub 2014 Jan 30.

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.

Keywords: cobalt incorporation; enzyme activation; nitrile hydratase; protein expression.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cobalt / metabolism
  • Enzyme Activation
  • Enzyme Activators / chemistry*
  • Enzyme Activators / metabolism*
  • Hydro-Lyases / chemistry*
  • Hydro-Lyases / genetics
  • Hydro-Lyases / metabolism*
  • Models, Molecular
  • Molecular Chaperones / chemistry*
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism*
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Pseudomonas putida / chemistry
  • Pseudomonas putida / enzymology*
  • Pseudomonas putida / genetics
  • Sequence Alignment

Substances

  • Bacterial Proteins
  • Enzyme Activators
  • Molecular Chaperones
  • Cobalt
  • Hydro-Lyases
  • nitrile hydratase