Inactivation of the bovine mitochondrial F1-ATPase with dicyclohexyl[14C]carbodiimide leads to the modification of a specific glutamic acid residue in the beta subunit.
When the bovine mitochondrial F1-ATPase is inactivated with dicyclohexyl[14C]carbodiimide and then gel-filtered, from 2 to 3 g atoms of 14C are incorporated/mol of enzyme. Prior inactivation of the enzyme by the modification of an essential tyrosine residue with 4-chloro-7-nitrobenzofurazan, a reaction that can be reversed by thiols, does not affect the irreversible inactivation of the ATPase by dicyclohexyl[14C]carbodiimide. During the large scale modification of the F1-ATPase by dicyclohexyl[14C]carbodiimide which led to 70% inactivation, 1.9 g atoms of 14C were incorporated/mol of enzyme. Isolation of the alpha, beta, and gamma subunits from this large scale inactivation revealed that the gram atoms of 14C bound per mol of each of the subunits was: alpha, 0.04; beta, 0.56; and gamma, 0.04. The majority of the radioactivity in a cyanogen bromide digest of the 14C-labeled beta subunit was isolated in a fragment that has the following amino acid sequence: Glu-Leu-Ile-Asn-Asn-Val-Ala-Lys-Ala-His-Gly-Gly-Tyr-Ser-Val-Phe-Ala-Gly-Val-Gly -Glu-Arg-Thr-Arg-Glu-Gly-Asn-Asp-Leu-Tyr-Glu*-His-Met; where Glu* represents the N gamma-glutamyl derivative of dicyclohexyl[14C]urea.
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002234:Carbodiimides / Q000494:pharmacology*; D002250:Carbon Radioisotopes; D002417:Cattle; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology*; D005971:Glutamates; D018698:Glutamic Acid; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology*; D010086:Oxidative Phosphorylation Coupling Factors / Q000037:antagonists & inhibitors*; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D006180:Proton-Translocating ATPases
D000596:Amino Acids; D002234:Carbodiimides; D002250:Carbon Radioisotopes; D005971:Glutamates; D046911:Macromolecular Substances; D010086:Oxidative Phosphorylation Coupling Factors; D010446:Peptide Fragments; D018698:Glutamic Acid; D004024:Dicyclohexylcarbodiimide; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases