Post-translational cleavage of recombinantly expressed nitrilase from Rhodococcus rhodochrous J1 yields a stable, active helical form

R. Ndoria Thuku, Brandon W. Weber, Arvind Varsani, B. Trevor Sewell

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Nitrilases convert nitriles to the corresponding carboxylic acids and ammonia. The nitrilase from Rhodococcus rhodochrous J1 is known to be inactive as a dimer but to become active on oligomerization. The recombinant enzyme undergoes post-translational cleavage at approximately residue 327, resulting in the formation of active, helical homo-oligomers. Determining the 3D structure of these helices using electron microscopy, followed by fitting the stain envelope with a model based on homology with other members of the nitrilase superfamily, enables the interacting surfaces to be identified. This also suggests that the reason for formation of the helices is related to the removal of steric hindrance arising from the 39 C-terminal amino acids from the wild-type protein. The helical form can be generated by expressing only residues 1-327.

Original languageEnglish (US)
Pages (from-to)2099-2108
Number of pages10
JournalFEBS Journal
Volume274
Issue number8
DOIs
StatePublished - Apr 2007
Externally publishedYes

Fingerprint

nitrilase
Rhodococcus
Oligomerization
Nitriles
Carboxylic Acids
Ammonia
Oligomers
Dimers
Electron microscopy
Electron Microscopy
Coloring Agents
Amino Acids
Enzymes
Proteins

Keywords

  • Electron microscopy
  • Helix
  • IHRSR
  • Nitrilase
  • Oligomeric form

ASJC Scopus subject areas

  • Biochemistry

Cite this

Post-translational cleavage of recombinantly expressed nitrilase from Rhodococcus rhodochrous J1 yields a stable, active helical form. / Thuku, R. Ndoria; Weber, Brandon W.; Varsani, Arvind; Sewell, B. Trevor.

In: FEBS Journal, Vol. 274, No. 8, 04.2007, p. 2099-2108.

Research output: Contribution to journalArticle

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