2D 1H NMR studies of oxidized 2(Fe 4S 4) ferredoxin from Clostridium pasteurianum

• Published in: FEBS letters 1991-09, Vol.289 (2), p.253-256
• Main Authors: Bertini, Ivano, Briganti, Fabrizio, Luchinat, Claudio, Messori, Luigi, Monnanni, Roberto, Scozzafava, Andrea, Vallini, Giovanni
• Format: Article
• Language: English
• Subjects: 2,2-dimethyl-2-silapentane-5-sulfonate; 2D NMR; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Clostridium - metabolism; Clostridium pasteurianum 2(Fe4S4) ferredoxin; correlation spectroscopy; COSY; CpFd; DSS; Ferredoxin; Ferredoxins - chemistry; Ferredoxins - metabolism; FID; free induction decay; Fundamental and applied biological sciences. Psychology; Hydrogen; Magnetic Resonance Spectroscopy - methods; Metalloprotein; Metalloproteins; mixing time; NOE; NOESY; nuclear Overhauser effect; nuclear Overhauser eftect spectroscopy; Other metalloproteins; Oxidation-Reduction; Protein Conformation; Proteins; recycle delay; Tron-sulfur cluster
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(91)81082-J

Oxidized ferredoxin from Clostridium pastetirianum, containing two Fe 4S 4 clusters, has been investigated using 2D 1H NMR spectroscopy at 600 MHz. 2D NMR experiments allowed complete assignment of the sixteen isotropically shifted signals corresponding to the β-CH 2 protons of the eight metal coordinated cysteines. Geminal connectivities of Cys β-CH 2 protons were identified through magnitude COSY experiments and confirmed through 2D NOESY experiments. A few additional signals could be assigned to the corresponding α-CH protons. The importance of 2D experiments to achieve firm assignments of isotropically shifted signals in paramagnetic metalloproteins is stressed.