title: Isolation, purification, and partial characterization of a membrane-bound Cl-/HCO3--activated ATPase complex from rat brain with sensitivity to GABAAergic ligands.
abstract: This study describes the isolation and purification of a protein complex with [Formula: see text]-ATPase activity and sensitivity to GABAAergic ligands from rat brain plasma membranes. The ATPase complex was enriched using size-exclusion, affinity, and ion-exchange chromatography. The fractions obtained at each purification step were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE), which revealed four subunits with molecular mass ∼48, 52, 56, and 59 kDa; these were retained at all stages of the purification process. Autoradiography revealed that the ∼52 and 56 kDa subunits could bind [3H]muscimol. The [Formula: see text]-ATPase activity of this enriched protein complex was regulated by GABAAergic ligands but was not sensitive to blockers of the NKCC or KCC cotransporters.
authors.affiliation: a Institute of General Pathology and Pathological Physiology , Department of General Pathophysiology , Moscow , Russia.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Menzikov
mesh_terms: D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D027321:Anion Transport Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001639:Bicarbonates / Q000378:metabolism*; D001921:Brain / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D002712:Chlorides / Q000378:metabolism*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008024:Ligands; D008297:Male; D011869:Radioligand Assay; D051381:Rats; D017208:Rats, Wistar; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
chemical_list: D027321:Anion Transport Proteins; D001639:Bicarbonates; D002712:Chlorides; D008024:Ligands; D005680:gamma-Aminobutyric Acid; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C031652:anion-sensitive ATPases
keywords: -ATPase; Autoradiography; Characterization of the enrichment -ATPase; Purification and characterization neuronal -ATPase; Purification of the neuronal -ATPase; characterization; chromatography; enrichment; purification; rat brain plasma membranes