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Vol. 273, Issue 2, 784-792, January 9, 1998

Bovine Adrenals Contain, in Addition to Ouabain, a Second Inhibitor of the Sodium Pump

Ralf Schneider, Victor Wray, Manfred Nimtz, Wolf Dieter Lehmannpar , Ulrike Kirch, Roberto Antolovic, and Wilhelm Schoner

From the Dagger  Institut für Biochemie und Endokrinologie, Justus-Liebig-Universität Giessen, Frankfurter Straße 100, D-35392 Giessen, Germany,  Gesellschaft für Biotechnologische Forschung, D-38124 Braunschweig, Germany, and par  Deutsches Krebsforschungszentrum, D-69120 Heidelberg, Germany

In the search for endogenous cardiac glycosides, two different inhibitors of the sodium pump have been isolated from bovine adrenals. Inhibitor A with a molecular mass of 600 Da and a UV maximum at 250 nm was purified from 16 kg of bovine adrenals. The pure substance (<1 ng) inhibited the sodium pump of human red blood cells with an affinity similar to that of ouabain, yet it cross-reacted with antibodies against the bufadienolide proscillaridin A but not against the cardenolide ouabain. Inhibitor A was slightly more hydrophilic than ouabain on RP-C18 high pressure liquid chromatography. Hence, it showed properties similar to the proscillaridin A immunoreactivity (Sich, B., Kirch, U., Tepel, M., Zideck, W., and Schoner, W. (1996) Hypertension 27, 1073-1078) that increased in humans with systolic blood pressure and pulse pressure.

Inhibitor B of the sodium pump with a molecular mass of 584 Da was purified 106-fold from 20 kg of bovine adrenals. It cross-reacted with antibodies against ouabain but not with antibodies against proscillaridin A and inhibited the sodium pump of human and rat red blood cells with the same affinity as ouabain. All other properties, such as the retention time in a C18-reversed phase chromatography, molecular mass determination by electrospray mass spectrometry and fragmentation pattern, and UV and 1H NMR spectroscopic data, were identical to ouabain. Hence, sodium pump inhibitor B from bovine adrenals is the cardenolide ouabain.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


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