Isolation and characterization of a sodium pump inhibitor from human plasma.
An endogenous sodium pump inhibitor has been purified from human plasma. The purification scheme involved large scale dialysis, extraction of lyophilized dialysate by methanol followed by preparative and semipreparative scale reverse-phase high-performance chromatography. A single peak of biologically active material was obtained enriched by a factor of greater than 10 billion. This material showed high chromatographic polarity, was inactivated by charring, strong acid, or alkali, and was resistant to short-term boiling. The purified material had a molecular weight between 300 and 900 g/mol and was insensitive to type I esterase and a variety of proteolytic enzymes. The purified factor inhibited the ouabain-sensitive uptake of 86Rb by human erythrocytes, binding of [3H]ouabain, and activity of dog kidney Na,K-adenosine triphosphatase (ATPase) with high affinity (less than 0.3 nM) in a time- and dose-dependent manner. Maximally effective concentrations of the digitalislike factor showed no effect on either human red blood cell Mg- or Ca-ATPase, rabbit muscle sarcoplasmic reticulum Ca-ATPase, or guinea pig stomach H,K-ATPase. The purified material is a highly potent selective inhibitor of the ion transport, receptor, and hydrolytic functions of the sodium pump. The characteristic properties of this substance suggest it may be a mammalian endogenous digitalis and may be similar to the sodium transport inhibitor detected in the plasma of volume-sensitive forms of experimental and human hypertension.
Hamlyn
J M
JM
Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland 21201.
Harris
D W
DW
Clark
M A
MA
Rogowski
A C
AC
White
R J
RJ
Ludens
J H
JH
D000328:Adult; D000368:Aged; D001798:Blood Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D002298:Cardenolides; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D004077:Digoxin; D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D012413:Rubidium / Q000097:blood; D012503:Saponins; D015222:Sodium Channels / Q000187:drug effects*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism
D001798:Blood Proteins; D002298:Cardenolides; D012503:Saponins; D015222:Sodium Channels; C034504:digoxin-like factors; D004077:Digoxin; D000254:Sodium-Potassium-Exchanging ATPase; D012413:Rubidium