Tuning of the FMN binding and oxido-reduction properties by neighboring side chains in Anabaena flavodoxin

Research output: Contribution to journalJournal articleResearchpeer-review

  • Susana Frago
  • Guillermina Goñi
  • Beatriz Herguedas
  • José Ramón Peregrina
  • Ana Serrano
  • Inmaculada Perez-Dorado
  • Molina, Rafael
  • Carlos Gómez-Moreno
  • Juan A Hermoso
  • Marta Martínez-Júlvez
  • Stephen G Mayhew
  • Milagros Medina

Contribution of three regions (phosphate-binding, 50's and 90's loops) of Anabaena apoflavodoxin to FMN binding and reduction potential was studied. Thr12 and Glu16 did not influence FMN redox properties, but Thr12 played a role in FMN binding. Replacement of Trp57 with Glu, Lys or Arg moderately shifted E(ox/sq) and E(sq/hq) and altered the energetic of the FMN redox states binding profile. Our data indicate that the side chain of position 57 does not modulate E(ox/sq) by aromatic stacking or solvent exclusion, but rather by influencing the relative strength of the H-bond between the N(5) of the flavin and the Asn58-Ile59 bond. A correlation was observed between the isoalloxazine increase in solvent accessibility and less negative E(sq/hq). Moreover, E(sq/hq) became less negative as positively charged residues were added near to the isoalloxazine. Ile59 and Ile92 were simultaneously mutated to Ala or Glu. These mutations impaired FMN binding, while shifting E(sq/hq) to less negative values and E(ox/sq) to more negative. These effects are discussed on the bases of the X-ray structures of some of the Fld mutants, suggesting that in Anabaena Fld the structural control of both electron transfer steps is much more subtle than in other Flds.

Original languageEnglish
JournalArchives of Biochemistry and Biophysics
Volume467
Issue number2
Pages (from-to)206-217
Number of pages12
ISSN0003-9861
DOIs
Publication statusPublished - 2007
Externally publishedYes

    Research areas

  • Anabaena/metabolism, Binding Sites, Enzyme Activation, Flavin Mononucleotide/chemistry, Flavodoxin/chemistry, Kinetics, Models, Chemical, Models, Molecular, Oxidation-Reduction, Protein Binding, Protein Structure, Tertiary

ID: 203019764