Characterization and N-terminal amino acid sequence of multiple ferredoxins in kidney and adrenal mitochondria.
Two separate ferredoxins that differ in molecular mass by about 1.5 kDa were isolated from both pig kidney and bovine adrenal mitochondria. The proteins had different biochemical and immunological properties and appeared to be distinct gene products. The smaller ferredoxin from pig kidney (renodoxin, Mr approximately equal to 13,500) was very similar to bovine adrenodoxin (Mr = 14,048). Both proteins had nearly identical N-terminal amino acid sequences and electron-transfer activities. However, renodoxin and adrenodoxin expressed distinct antigenic determinants, although they were immunologically cross-reactive. The larger kidney (approximately equal to 15-kDa) and adrenal (approximately equal to 15.3-kDa) ferredoxins were biochemically similar to each other but they had lower specific activities and their N-terminal sequences were different when compared to renodoxin and adrenodoxin. Each of the four ferredoxins had a visible absorption spectrum characteristic of a [2Fe-2S] chromophore. But in addition, the larger ferredoxins displayed a prominent A276 peak due to their higher tyrosine content and the presence of tryptophan, which is absent in adrenodoxin and renodoxin. Consequently, the larger ferredoxins were termed Trp-ferredoxin. Using antibody to pig kidney Trp-ferredoxin, the larger adrenal and kidney ferredoxins were found to be very similar immunologically; however, the Trp-containing and adrenodoxin-type ferredoxins did not cross-react in immunoblot analysis. Nevertheless, it was shown from competition ELISA and activity-inhibition analysis that the two ferredoxin types had limited common antigenic determinants. Trp-ferredoxin was the major iron-sulfur protein in kidney whereas adrenodoxin was the dominant molecular form in adrenal gland.
Driscoll
W J
WJ
Department of Biochemistry, University of New Mexico School of Medicine, Albuquerque 87131.
Omdahl
J L
JL
D000311:Adrenal Glands / Q000032:analysis*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D003574:Cytochrome c Group / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005288:Ferredoxins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D015151:Immunoblotting; D007668:Kidney / Q000032:analysis*; D008928:Mitochondria / Q000032:analysis*; D013552:Swine
D003574:Cytochrome c Group; D005288:Ferredoxins; D003577:Cytochrome P-450 Enzyme System