Purification and characterization of a membrane-bound ATPase from Acetabularia cliftonii that corresponds to a Cl(-)-translocating ATPase in Acetabularia acetabulum.
A Mg(2+)-ATPase was solubilized from membranes of Acetabularia cliftonii using nonanoyl-N-methylgluconamide and purified by ion-exchange and gel permeation chromatography. One active ATPase fraction after Mono Q chromatography had a specific activity of 10 units/mg of protein. Judged from subunit composition [54 (a), 50 (b) with a fainter band around 40 kDa], catalytic properties, and N-terminal amino acid sequence of the b subunit, the isolated enzyme was comparable to the Cl(-)-ATPase of Acetabularia acetabulum. Immunological characterization of both subunits showed significant similarity to the F type of ATPase. Cl(-)-transport activity was observed by reconstitution studies into liposomes.
Moritani
C
C
Faculty of Engineering Sciences, Okayama University, Japan.
Ohhashi
T
T
Satoh
S
S
Oesterhelt
D
D
Ikeda
M
M
D000076:Acetabularia / Q000201:enzymology*; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D001692:Biological Transport; D002712:Chlorides / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000378:metabolism; D013379:Substrate Specificity
D002712:Chlorides; D008565:Membrane Proteins; D009711:Nucleotides; D000251:Adenosine Triphosphatases