Secondary and tertiary structure characteristics of Megasphaera elsdenii flavodoxin in the reduced state as determined by two-dimensional 1H NMR.
The secondary structure of two-electron-reduced Megasphaera elsdenii flavodoxin has been determined by visual, qualitative inspection of the sequential connectivities involving C alpha H, C beta H and NH protons observed in NOESY (two-dimensional nuclear Overhauser enhancement spectroscopy) spectra. Results from an amide proton exchange experiment were used to confirm the secondary structure assignment and to demonstrate the compactness and stability of the protein. After the secondary structure elements were established, the global fold of the protein and the flavin binding site have been determined using nonsequential interresidual NOE connectivities as primary source of information. The secondary structure and the global fold of M. elsdenii and Clostridium MP flavodoxin appeared to be very similar, differences are observed however. M. elsdenii flavodoxin consists of a central parallel beta-sheet including five strands surrounded on both sides by a pair of alpha-helices.
van Mierlo
C P
CP
Department of Biochemistry, Agricultural University, Wageningen, The Netherlands.
Müller
F
F
Vervoort
J
J
D000577:Amides; D000595:Amino Acid Sequence; D001419:Bacteria / Q000032:analysis*; D003013:Clostridium / Q000032:analysis; D005418:Flavodoxin / Q000032:analysis*; D005420:Flavoproteins / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011522:Protons
D000577:Amides; D005418:Flavodoxin; D005420:Flavoproteins; D011522:Protons