Molecular cloning and immunological characterization of porcine kidney ferredoxin.
Porcine renodoxon is a kidney mitochondrial iron-sulfur protein (ISP) that functions to transfer electron to cytochromes P450 of the vitamin D pathway. A full-length cDNA clone to porcine renodoxin was isolated in the current investigation and used to study the protein's primary structure and immunological properties. The cysteine ligands for the iron-sulfur center, and the surface protein-binding and phosphorylation sites occupied identical positions in both porcine renodoxin and bovine adrenodoxin. Furthermore, porcine renodoxin was functionally indistinguishable from bovine adrenodoxin and the mature forms of both proteins had the same encoded length and shared approximately 91% sequence similarity. A synthetic peptide to the surface protein-binding region was used to demonstrate the antigenicity of the domain in both the porcine and the bovine ISPs. However, porcine renodoxin displayed only limited immunological identity to other regions of bovine adrenodoxin as measured by competitive enzyme-linked immunosorbent assay. Part of this immunological distinction was attributed to the COOH-terminal processing of porcine renodoxin, an action which negated expression of a COOH-terminal antigenic site that is present in bovine adrenodoxin. Other antigenic differences were linked to charged-residue substitutions that were located in predicted surface domains. The highest frequency of surface-residue substitutions in ferredoxin proteins was predicted for porcine renodoxin, which could provide a basis for understanding why the pig protein appears more antigenically divergent than other ferredoxins.
Omdahl
J L
JL
Department of Biochemistry, University of New Mexico School of Medicine, Albuquerque 87131.
Wilson
K
K
Swerdlow
H
H
Driscoll
W J
WJ
D000325:Adrenodoxin / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D003429:Cross Reactions; D004247:DNA / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D005288:Ferredoxins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D007668:Kidney / Q000737:chemistry*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D013552:Swine
D000954:Antigens, Surface; D005288:Ferredoxins; D000325:Adrenodoxin; D004247:DNA