title: Unusual lack of internal mobility and fast overall tumbling in oxidized flavodoxin from Anacystis nidulans.
abstract: Anacystis nidulans flavodoxin, an electron-transfer protein containing a flavin mononucleotide (FMN) molecule as its prosthetic group, has a redox potential for the oxidized/semiquinone equilibrium close to that of free flavin. Whereas the redox potential for the semiquinone/hydroquinone equilibrium is more negative. To gain an understanding of the contribution of mobility to redox potential modulation, we studied the backbone mobility of the oxidized A. nidulans flavodoxin at pH 6.6, 303 K by 15N NMR relaxation measurements. The spin-lattice relaxation rate constants (RN(Nz)=1/T1), spin-spin relaxation rate constants (RN(Nx,y)=1/T2) and 1H-15N nuclear Overhauser effects (NOE) were obtained for 143 of the 166 protonated backbone 15N nuclei and for the FMN N3 nucleus without ambiguity. The 15N T1, T2 and NOE data were analyzed by reduced spectral density mapping, and the so-called model-free approach. In contrast to most other proteins studied with 15N relaxation experiments, we found an almost complete absence of internal mobility. The overall correlation time of this 169-residue flavodoxin (>19 kDa) is significantly shorter (7.4 to 7.8 ns) than that of other proteins of this size, suggesting that the absence of internal mobility is correlated with faster overall rotational diffusion. The uniformity of the motional parameters along the backbone is in strong contrast to the crystallographic B-factors, which vary significantly along the sequence in this and other flavodoxins. The NMR relaxation parameters are primarily sensitive to rotational diffusive motions of the N-H bond vectors, while the crystallographic B-factors would be sensitive to translational internal motions as well. However, the large B-factors in this protein may originate from crystal packing and crystal lattice disorder. The relatively fast overall tumbling results in sharp NMR resonances. Hence, much larger proteins with such favorable dynamic behavior could be excellent candidates for studies by NMR.
authors.affiliation: Committee on Higher Degrees in Biophysics, Harvard University, 240 Longwood Avenue, Boston, MA 02115, USA.
authors.forename: P
authors.initials: P
authors.lastname: Zhang
authors.forename: K T
authors.initials: KT
authors.lastname: Dayie
authors.forename: G
authors.initials: G
authors.lastname: Wagner
mesh_terms: D003198:Computer Simulation; D000458:Cyanobacteria / Q000737:chemistry*; D005486:Flavin Mononucleotide / Q000737:chemistry*; D005418:Flavodoxin / Q000737:chemistry* / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008962:Models, Theoretical; D009038:Motion; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013045:Species Specificity
chemical_list: D005418:Flavodoxin; D011994:Recombinant Proteins; D005486:Flavin Mononucleotide