Ultrastructure of membrane-bound Na, K-ATPase after extensive tryptic digestion.
Membrane-bound Na, K-ATPase was digested with trypsin in the presence of Rb+ to form the stable 19-kDa and smaller fragments of the alpha-chain known to preserve occlusion of Rb+ (K+) or Na+. The trypsinized membranes obtained from pig kidney and shark rectal gland were analyzed by electron microscopy. Tryptic digestion preserved general membrane structure but removed both the surface particles observed by negative staining and the protruding cytoplasmic portion of the alpha-subunit identified in thin sections. However, intramembrane particles defined by freeze-fracture were preserved after trypsinization suggesting that the remaining membrane spanning protein fragments retain the native structure within the lipid bilayer after proteolysis.
Department of Cell Biology, University of Aarhus, Denmark.
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Maunsbach
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Esmann
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000201:enzymology; D008854:Microscopy, Electron; D010446:Peptide Fragments / Q000378:metabolism; D012007:Rectum / Q000201:enzymology; D012491:Salt Gland / Q000201:enzymology; D012754:Sharks; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism / Q000648:ultrastructure*; D013552:Swine; D014357:Trypsin / Q000378:metabolism
D010446:Peptide Fragments; D014357:Trypsin; D000254:Sodium-Potassium-Exchanging ATPase