Tuning of the FMN Binding and Oxido-Reduction Properties by Neighboring Side Chains in Anabaena Flavodoxin

Frago, Susana (Universidad de Zaragoza) ; Goñi, Guillermina (Universidad de Zaragoza) ; Herguedas, Beatriz ; Peregrina, Jose Ramon ; Serrano , Ana (Universidad de Zaragoza) ; Perez-Dorado, Inmaculada ; Molina, Rafael ; Gomez-Moreno, Carlos ; Hermoso, Juan A. ; Martinez-Julvez, Marta (Universidad de Zaragoza) ; Mayhew, Stephen G. ; Medina, Milagros (Universidad de Zaragoza)

Resumen: 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 Eox/sq and Esq/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 Eox/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 Esq/hq. Moreover, Esq/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 Esq/hq to less negative values and Eox/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.

Idioma: Inglés

DOI: 10.1016/j.abb.2007.08.024

Año: 2007

Publicado en: ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 467, 2 (2007), 206-217
ISSN: 0003-9861

Factor impacto JCR: 2.578 (2007)

Categ. JCR: BIOPHYSICS rank: 32 / 67 = 0.478 (2007) - Q2 - T2
Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 131 / 262 = 0.5 (2007) - Q2 - T2

Tipo y forma: Article (PostPrint)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)