Localization and identification of covalently bound flavoproteins in rat liver mitochondria by prelabeling of their flavin moiety.
The covalently bound flavoproteins in rat liver mitochondria were prelabeled by injecting [14C]riboflavin into a rat, then liver mitochondria were obtained and further labeled with [3H]pargyline, a suicide inhibitor of monoamine oxidase. When the mitochondria were subjected to osmotic lysis, two covalently bound flavoproteins having molecular weights of 110,000 and 94,000 were found in the supernatant. These proteins were identified as sarcosine dehydrogenases. Upon treatment of the membranous fraction with 1% Triton X-100, succinate dehydrogenase with a molecular weight of 70,000 was found in the soluble fraction, while two well-separated proteins doubly-labeled with 14C and 3H were found in the insoluble fraction. Their molecular weights were 61,000 and 57,000. By isoelectric focusing, two 3H peaks were observed with pI values of 8.3 and 8.4. The former corresponded to the 61,000-dalton protein, and the latter, to the 57,000 one. From the data obtained by using selective inhibitors, deprenyl and clorgyline, the [3H]pargyline-binding proteins with molecular weights of 61,000 and 57,000 were assigned to proteins of monoamine oxidases of type A and type B, respectively.
D000818:Animals; D001596:Benzylamines / Q000378:metabolism; D002250:Carbon Radioisotopes; D003010:Clorgyline / Q000494:pharmacology; D005420:Flavoproteins / Q000032:analysis*; D008297:Male; D008930:Mitochondria, Liver / Q000378:metabolism*; D008970:Molecular Weight; D008995:Monoamine Oxidase / Q000032:analysis*; D010089:Oxidoreductases, N-Demethylating / Q000032:analysis*; D010293:Pargyline / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D012256:Riboflavin / Q000378:metabolism; D050892:Sarcosine Dehydrogenase; D012642:Selegiline / Q000494:pharmacology; D012701:Serotonin / Q000378:metabolism; D014316:Tritium; D014439:Tyramine / Q000378:metabolism
D001596:Benzylamines; D002250:Carbon Radioisotopes; D005420:Flavoproteins; D014316:Tritium; D012642:Selegiline; D012701:Serotonin; D010293:Pargyline; C030796:benzylamine; D008995:Monoamine Oxidase; D010089:Oxidoreductases, N-Demethylating; D050892:Sarcosine Dehydrogenase; C494397:Sardh protein, rat; D003010:Clorgyline; D012256:Riboflavin; D014439:Tyramine