Fo portion of Escherichia coli ATP synthase: orientation of subunit c in the membrane
- 25 August 1987
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 26 (17), 5486-5492
- https://doi.org/10.1021/bi00391a041
Abstract
Incubation of right-side-out oriented membrane vesicles of Escherichia coli with tetranitromethane resulted in the nitration of tyrosine residues (Tyr-10 and Tyr-73) of subunit c from the ATP synthase. Cleavage of the protein with cyanogen bromide and separation of the resulting fragments, especially of the tyrosine-containing peptides, clearly demonstrated that the distribution of the nitro groups is similar at any time and at any pH value chosen for the analysis. Furthermore, the percentage of 3-nitrotyrosine present in the two peptide fragments was in good agreement with that obtained for the intact polypeptide chain. While the modification of the tyrosine residues in subunit c with the lipophilic tetranitromethane is independent of the orientation of the membranes vesicles, the subsequent partial conversion of the 3-nitrotyrosine to the amino form only occurred when membrane vesicles with right-side-out orientation were treated with the ionic, water-soluble sodium dithionite, which at certain concentrations cannot penetrate biological membranes. Cleavage of subunit c isolated from nitrated and subsequently reduced membrane vesicles and separation of the resulting fragments by high-pressure liquid chromatography showed that the 3-nitrotyrosine in the Tyr-73-containing peptides has been completely reduced, while the nitro group in peptides containing Tyr-10 remained nearly unaffected.This publication has 21 references indexed in Scilit:
- Membrane integration and function of the three F0 subunits of the ATP synthase of Escherichia coli K12.The EMBO Journal, 1983
- Stoichiometry of subunits in the H+-ATPase complex of Escherichia coli.Journal of Biological Chemistry, 1982
- Identification of Amino‐Acid Substitutions in the Proteolipid Subunit of the ATP Synthase from Dicyclohexylcarbodiimide‐Resistant Mutants of Escherichia coliEuropean Journal of Biochemistry, 1980
- N,N'-dicyclohexylcarbodiimide binds specifically to a single glutamyl residue of the proteolipid subunit of the mitochondrial adenosinetriphosphatases from Neurospora crassa and Saccharomyces cerevisiae.Proceedings of the National Academy of Sciences, 1980
- Purification of the carbodiimide-reactive protein component of the ATP energy-transducing system of Escherichia coli.Journal of Biological Chemistry, 1976
- [44] Nitration with tetranitromethaneMethods in Enzymology, 1972
- [13] Bacterial MembranesPublished by Elsevier ,1971
- Tetranitromethane. A Reagent for the Nitration of Tyrosyl Residues in Proteins*Biochemistry, 1966
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951
- MUTANTS OF ESCHERICHIA COLI REQUIRING METHIONINE OR VITAMIN B 12Journal of Bacteriology, 1950