PURIFICATION AND BIOCHEMICAL-CHARACTERIZATION OF A PUTATIVE [6FE-6S] PRISMANE-CLUSTER-CONTAINING PROTEIN FROM DESULFOVIBRIO-VULGARIS (HILDENBOROUGH)

A novel iron-sulfur protein has been isolated from the sulfate-reducing bacterium Desulfovibrio vulgaris (Hildenborough). It is a stable monomeric protein, which has a molecular mass of 52 kDa, as determined by sedimentation-equilibrium centrifugation. Analysis of the metal and acid-labile sulfur content of the protein revealed the presence of 6.3 +/- 0.4 Fe/polypeptide and 6.2 +/- 0.7 S2-/polypeptide. Non-iron transition metals, heme, flavin and selenium were absent. Combining these data with the observation of a very anisotropic S = 1/2 [6Fe-6S]3+ prismane-like EPR signal in the dithionite-reduced protein, we believe that we have encountered the first example of a prismane-cluster-containing protein. The prismane protein has a slightly acidic amino acid composition and isoelectric point (pI = 4.9). The ultraviolet/visible spectrum is relatively featureless (epsilon-280 = 81 mM-1.cm-1, epsilon-400 = 25 mM-1.cm-1, epsilon-400,red = 14 mM-1.cm-1). The shape of the protein is approximately globular (S20,w = 4.18 S). The N-terminal amino acid sequence is MF(S)/(C)FQ(S)/C QETAKNTG. Polyclonal antibodies against the protein were raised. Cytoplasmic localization was inferred from subcellular fractionation studies. Cross-reactivity of antibodies against this protein indicated the occurrence of a similar protein in D. vulgaris (Monticello) and Desulfovibrio desulfuricans (ATCC 27774). We have not yet identified a physiological function for the prismane protein despite trials for some relevant enzyme activities.