Revealing of proteins interacting with Na,K-ATPase.
Proteins interacting with alpha 1 beta 1-type of Na,K-ATPase were revealed in pig kidney outer medulla and duck salt glands using three different methods (immunoprecipitation, protein overlay, and chemical cross-linking). Immunoprecipitation was performed after solubilization of protein homogenate with Triton X-100 so that both membrane and cytosol proteins bound to Na,K-ATPase could be revealed. Two other methods were used to study the interaction of cytosol proteins with purified Na,K-ATPase. The sets of proteins revealed by each method in outer medulla of pig kidney were different. Proteins interacting with Na,K-ATPase that have molecular masses 10, 15, 70, 75, 105, 120, and 190 kD were found using the immunoprecipitation method. The chemical cross-linking method revealed proteins with molecular masses 25, 35, 40, 58, 68-70, and 86-88 kD. The protein overlay method revealed in the same tissue proteins with molecular masses 38, 42, 43, 60, 62, 66, 70, and 94 kD.
Akimova
O A
OA
Department of Biochemistry, Faculty of Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
Dolgova
N V
NV
Mast
N V
NV
Rubtsov
A M
AM
Lopina
O D
OD
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D003432:Cross-Linking Reagents; D003600:Cytosol; D004372:Ducks; D007679:Kidney Medulla / Q000166:cytology / Q000378:metabolism; D008970:Molecular Weight; D011233:Precipitin Tests; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012491:Salt Gland / Q000166:cytology / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D013552:Swine
D003432:Cross-Linking Reagents; D011506:Proteins; D000254:Sodium-Potassium-Exchanging ATPase