Cell-free synthesis and characterization of human adrenocortical pro-adrenodoxin.
Poly(A+)-RNAs were extracted from human hyperplasic adrenocortical tissue and translated in a wheat germ cell-free system in the presence of [35S]-methionine. Labeled immuno-reactive adrenodoxin (ADX)-like material was immunoisolated and examined following mono and bi-dimensional electrophoretic analysis. Bovine mRNA translation products were analysed under similar conditions. While it was confirmed that bovine ADX was synthesized as a precursor of Mr 21 kDa, human pro-ADX was characterized for the first time as a somewhat larger moiety (24 kDa). On the other hand, both human and bovine mature mitochondrial ADX showed a Mr of 12 kDa. Electrophoretic study disclosed that the human, as well as the bovine pro-ADX could be resolved into several components differing by their pHi (6.5 and 6.9 for h-proADX and 5.9, 6.1 and 6.2 for b-proADX, respectively). This molecular heterogeneity might be explained by discrete disparity in the pro-adrenodoxin amino acid contents.
Pascal
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Monnier
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Chambaz
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EM
D000302:Adrenal Cortex / Q000378:metabolism*; D000325:Adrenodoxin / Q000096:biosynthesis*; D000818:Animals; D002417:Cattle; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D008715:Methionine / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis*; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism
D011498:Protein Precursors; D012333:RNA, Messenger; D000325:Adrenodoxin; D011061:Poly A; D012313:RNA; D008715:Methionine