Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri.
AUTOR(ES)
Inatomi, K
RESUMO
Membrane-bound ATPase was found in membranes of the archaebacterium Methanosarcina barkeri. The ATPase activity required divalent cations, Mg2+ or Mn2+, and maximum activity was obtained at pH 5.2. The activity was specifically stimulated by HSO3- with a shift of optimal pH to 5.8, and N,N'-dicyclohexylcarbodiimide inhibited ATP hydrolysis. The enzyme could be solubilized from membranes by incubation in 1 mM Tris-maleate buffer (pH 6.9) containing 0.5 mM EDTA. The solubilized ATPase was purified by DEAE-Sepharose and Sephacryl S-300 chromatography. The molecular weight of the purified enzyme was estimated to be 420,000 by gel filtration through Sephacryl S-300. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate revealed two classes of subunit, Mr 62,000 (alpha) and 49,000 (beta) associated in the molar ratio 1:1. These results suggest that the ATPase of M. barkeri is similar to the F0F1 type ATPase found in many eubacteria.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=215949Documentos Relacionados
- Identification of a vanadate-sensitive, membrane-bound ATPase in the archaebacterium Methanococcus voltae.
- Characterization and solubilization of the membrane-bound ATPase of Mycoplasma gallisepticum.
- Characterization and purification of carbon monoxide dehydrogenase from Methanosarcina barkeri.
- Purification and characterization of the membrane-bound ferrochelatase from Spirillum itersonii.
- Purification and Characterization of a Membrane-Bound Hydrogenase from the Hyperthermophilic Archaeon Pyrococcus furiosus