A single-residue mutation destabilizes Vibrio harveyi flavin reductase FRP dimer

Navneet Jawanda, Jerry Ebalunode, Alexey Gribenko, James Briggs, James Lee, Shiao Chun Tu

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Our earlier studies have shown that the Vibrio harveyi flavin reductase FRP undergoes a monomer-dimer equilibrium, and luciferase forms a functional complex with the FRP monomer but not significantly with the dimer. This work is aimed at further investigating the nature and regulation of FRP subunit interactions by computation and site-directed mutagenesis approaches. In silico mutations of a number of residues were performed, and energetic analyses led us to target residue E99, which interacts directly with R113 and R225 from the second subunit of the FRP homodimer, for detailed investigation. E99 was found non-essential to the binding of either the FMN cofactor or the substrates. However, in comparison with the native enzyme, the E99K variant was shown to have an enhanced subunit dissociation as evident from a 44-fold higher Kd for the monomer-dimer equilibrium. The critical role of E99 in the formation of the FRP dimer has thus been demonstrated.

Original languageEnglish (US)
Pages (from-to)51-57
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume472
Issue number1
DOIs
StatePublished - Apr 1 2008

Fingerprint

sudan III
Flavin Mononucleotide
Vibrio
Site-Directed Mutagenesis
Luciferases
Dimers
Computer Simulation
Oxidoreductases
Mutation
Monomers
Enzymes
Mutagenesis
4,6-dinitro-o-cresol
Substrates

Keywords

  • bacterial
  • Dimmer dissociation
  • Flavin reductase
  • Luciferase
  • Monomer-dimer equilibrium
  • Subunit interaction
  • Vibrio harveyi

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A single-residue mutation destabilizes Vibrio harveyi flavin reductase FRP dimer. / Jawanda, Navneet; Ebalunode, Jerry; Gribenko, Alexey; Briggs, James; Lee, James; Tu, Shiao Chun.

In: Archives of Biochemistry and Biophysics, Vol. 472, No. 1, 01.04.2008, p. 51-57.

Research output: Contribution to journalArticle

Jawanda, Navneet ; Ebalunode, Jerry ; Gribenko, Alexey ; Briggs, James ; Lee, James ; Tu, Shiao Chun. / A single-residue mutation destabilizes Vibrio harveyi flavin reductase FRP dimer. In: Archives of Biochemistry and Biophysics. 2008 ; Vol. 472, No. 1. pp. 51-57.
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AU - Tu, Shiao Chun

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