The proteolipid of the A(1)A(0) ATP synthase from Methanococcus jannaschii has six predicted transmembrane helices but only two proton-translocating carboxyl groups
Autor(en): | Ruppert, C Kavermann, H Wimmers, S Schmid, R Kellermann, J Lottspeich, F Huber, H Stetter, KO Muller, V |
Stichwörter: | ARCHAEON; Biochemistry & Molecular Biology; COENZYME-M METHYLTRANSFERASE; ESCHERICHIA-COLI; EVOLUTION; EXPRESSION; F-0 SECTOR; METHANOSARCINA-MAZEI GO1; ORGANIZATION; STOICHIOMETRY; STRAIN GO1 | Erscheinungsdatum: | 1999 | Herausgeber: | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC | Journal: | JOURNAL OF BIOLOGICAL CHEMISTRY | Volumen: | 274 | Ausgabe: | 36 | Startseite: | 25281 | Seitenende: | 25284 | Zusammenfassung: | The proteolipid, a hydrophobic ATPase subunit essential for ion translocation, was purified from membranes of Methanococcus jannaschii by chloroform/methanol extraction and gel chromatography and was studied using molecular and biochemical techniques. Its apparent molecular mass as determined in SDS-polyacrylamide gel electrophoresis varied considerably with the conditions applied. The N-terminal sequence analysis made it possible to define the open reading frame and revealed that the gene is a triplication of the gene present in bacteria. In some of the proteolipids, the N-terminal methionine is excised. Consequently, two forms with molecular masses of 21,316 and 21,183 Da were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The molecular and biochemical data gave clear evidence that the mature proteolipid from M. jannaschii is a triplication of the 8-kDa proteolipid present in bacterial F1F0 ATPases and most archaeal A(1)A(0) ATPases, Moreover, the triplicated form lacks a proton-translocating carboxyl group in the first of three pairs of transmembrane helices, This finding puts in question the current view of the evolution of H+ ATPases and has important mechanistic consequences for the structure and function of H+ ATPases in general. |
ISSN: | 00219258 | DOI: | 10.1074/jbc.274.36.25281 |
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