2-DIMENSIONAL H-1-NMR STUDIES ON DESULFOVIBRIO-GIGAS FERREDOXINS - ASSIGNMENT OF THE IRON-SULFUR CLUSTER CYSTEINYL LIGAND PROTONS

被引:28
作者
MACEDO, AL
PALMA, PN
MOURA, I
LEGALL, J
WRAY, V
MOURA, JJG
机构
[1] UNIV NOVA LISBOA,FAC CIENCIAS & TECNOL,DEPT QUIM,P-2825 MONTE DE CAPARICA,PORTUGAL
[2] UNIV NOVA LISBOA,INST TECNOL QUIM & BIOL,P-2780 OEIRAS,PORTUGAL
[3] UNIV GEORGIA,DEPT BIOCHEM,ATHENS,GA 30602
[4] GESELL BIOTECHNOL FORSCH MBH,MOLEK STRUKTURFORSCH ABT,W-3300 BRAUNSCHWEIG,GERMANY
关键词
NMR; H-1-NMR; NOESY; TOCSY; DESULFOVIBRIO SP; FERREDOXINS; 3FE-4S] AND [4FE-4S; CLUSTERS; 2D NMR; IRON-SULFUR CLUSTERS;
D O I
10.1002/mrc.1260311313
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
1D and 2D H-1 NMR studies are reported on the oxidized and reduced [4Fe-4S] cluster of Desulfovibrio gigas ferredoxin I (FdI). Several low-field contact shifted resonances (fast relaxing) are assigned to beta-CH2 and alpha-CH coordinated cysteinyl residues. NOESY patterns (supported by 1D NOE experiments) resolves four pairs of geminal beta-CH2 protons at low-field. The cluster ligands are assigned non-specifically to Cys8, Cys11, Cys14 and Cys50, based on the X-ray structural analysis available for the oligomeric form, FdII, that contains a single [3Fe-4S] cluster. It was indicated in this case that Cys11 is not bound to the trinuclear cluster but is tilted towards the solvent. The presence of four pairs of geminal beta-CH2 protons for FdI unambiguously proves the occupancy of the fourth site of the [3Fe-4S] complex and implies the coordination of the Cys11 at the cluster. Analysis of the oxidized form of FdII, using the same methodology as described for FdI, supports the presence of three cysteinyl ligands in the [3Fe-4S] core. Further, the combined use of the X-ray coordinates enables the specific assignment of the three cysteinyl ligands of the cluster, extending a previous assignment of Cys50. In addition, very broad resonances were detected for the reduced form of FdII in the low-field region around 200 ppm and in the high field region around -80 ppm.
引用
收藏
页码:S59 / S67
页数:9
相关论文
共 60 条
  • [41] MACURA SR, 1980, MOL PHYS, V40, P95
  • [42] MARKLEY JL, 1986, IRON SULFUR PROTEIN, P167
  • [43] MORENO C, IN PRESS J INORG BIO
  • [44] MORENO C, 1991, 3RD INT C MOL BIOL H, P156
  • [45] EVIDENCE FOR THE FORMATION OF A COFE3S4 CLUSTER IN DESULFOVIBRIO-GIGAS FERREDOXIN-H
    MOURA, I
    MOURA, JJG
    MUNCK, E
    PAPAEFTHYMIOU, V
    LEGALL, J
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1986, 108 (02) : 349 - 351
  • [46] NMR CHARACTERIZATION OF 3 FORMS OF FERREDOXIN FROM DESULFOVIBRIO-GIGAS, A SULFATE REDUCER
    MOURA, JJG
    XAVIER, AV
    BRUSCHI, M
    LEGALL, J
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1977, 459 (02) : 278 - 289
  • [47] MOURA JJG, 1982, J BIOL CHEM, V257, P6259
  • [48] MOURA JJG, 1977, FEBS LETT, V89, P177
  • [49] MUNCK E, 1988, ACS SYM SER, V372, P302
  • [50] NMR-STUDIES ON FERREDOXINS .1. CLASSIFICATION OF IRON-SULFUR CORES IN FERREDOXINS BY H-1 NUCLEAR MAGNETIC-RESONANCE SPECTROSCOPY
    NAGAYAMA, K
    OZAKI, Y
    KYOGOKU, Y
    HASE, T
    MATSUBARA, H
    [J]. JOURNAL OF BIOCHEMISTRY, 1983, 94 (03) : 893 - 902