Origin of the gamma polypeptide of the Na+/K+-ATPase.
The Na+/K+-ATPase purified from lamb kidney contains a gamma polypeptide fraction which is a collection of fragments derived from the alpha and beta polypeptides of the enzyme. This fraction has the solubility characteristics of a proteolipid and was isolated either by high performance liquid chromatography (size exclusion chromatography) in 1% sodium dodecyl sulfate or by sequential organic extraction of purified lamb kidney Na+/K+-ATPase. Formation of gamma polypeptide(s) from detergent solubilized holoenzyme was accelerated by sulfhydryl containing reagents and was unaffected by addition of inhibitors of proteolytic enzymes. Treatment of the holoenzyme with the photoaffinity reagent N-(2-nitro-4-azidophenyl)[3H]ouabain ([3H]NAP-ouabain) labeled the alpha polypeptide and the gamma polypeptide fraction but not the beta polypeptide. Amino acid sequence analysis of one gamma polypeptide preparation revealed homology of one component of this fraction with the N-terminus of the beta subunit of the Na+/K+-ATPase. Amino acid analysis of two preparations of proteolipid showed similar amino acid compositions with a peptide derived from the alpha subunit. The insolubility and complexity of the gamma polypeptide(s)/proteolipid fraction appears to preclude a conclusive sequence analysis of all components of this fraction.
Harris
W E
WE
Neurochemistry Laboratory, VA Medical Center, Seattle, WA 98108.
Stahl
W L
WL
D000345:Affinity Labels / Q000494:pharmacology; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001386:Azides / Q000494:pharmacology; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D007668:Kidney / Q000201:enzymology; D008623:Mercaptoethanol / Q000494:pharmacology; D010042:Ouabain / Q000031:analogs & derivatives / Q000494:pharmacology; D010446:Peptide Fragments / Q000032:analysis*; D011510:Proteolipids / Q000032:analysis; D012756:Sheep; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis*
D000345:Affinity Labels; D000596:Amino Acids; D001386:Azides; D010446:Peptide Fragments; D011510:Proteolipids; D010042:Ouabain; D008623:Mercaptoethanol; C032780:NAP-ouabain; D000254:Sodium-Potassium-Exchanging ATPase