Three-dimensional correlated NMR study of Megasphaera elsdenii flavodoxin in the oxidized state.
The value of a three-dimensional (3D) non-selective total correlation/nuclear Overhauser enhancement spectroscopy (TOCSY-NOESY) spectrum for making sequential resonance assignments in proteins is demonstrated using the relatively large Megasphaera elsdenii flavodoxin (molecular mass 15 kDa) in the oxidized state. An easy and concise method for the analysis of 3D-NMR spectra and a strategy for the resonance assignment of 3D-NMR protein spectra is introduced. In this context, non-selective TOCSY-NOESY is compared with selective TOCSY-NOESY and non-selective NOESY-TOCSY. Sequential assignments in various secondary structure elements of flavodoxin are made using the method of analysis introduced. NOEs not previously identified in 2D-NMR spectra due to resonance overlap are found in the 3D Clean-TOCSY-NOESY spectrum. Also additional side-chain assignments could be made.
Wijmenga
S S
SS
National HF-NMR Facility, Faculty of Science, University of Nijmegen, The Netherlands.
van Mierlo
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CP
D000595:Amino Acid Sequence; D005418:Flavodoxin / Q000737:chemistry* / Q000302:isolation & purification; D006089:Gram-Negative Anaerobic Bacteria / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation
D005418:Flavodoxin