A novel endogenous sodium-pump inhibitor in pig urine: purification and comparison with the inhibitor purified from bovine adrenal glands.
To clarify the chemical nature and pathophysiological significance of an endogenous, specific Na-pump inhibitor, extracts of various tissues have been examined. The activities were extracted by an Amberlite XAD-2 adsorbent and fractionated by high-performance liquid chromatography (HPLC) on an ODS column. Among many tissues and fractions tested, the extract of pig urine has been found to contain a large amount of 86Rb uptake inhibitory activities. The strongest inhibitory activity for 86Rb influx into erythrocytes also exhibited a cross-reactivity with anti-ouabain antibodies. This activity has been purified to homogeneity using five steps of reverse-phase HPLC. Although most of the dose-response curves for this purified Na-pump inhibitor, designated as uroxin, in the various assay systems paralleled those of ouabain and the inhibitor purified from bovine adrenal glands (designated as adrexin C), the cross-reactivity curve with anti-ouabain antibodies did not. The retention time of uroxin on an ODS HPLC column was also different from that of digoxin, ouabain, or adrexin C. 1H nuclear magnetic resonance spectroscopic study suggests that uroxin is a novel Na-pump inhibitor that is structurally different from any of the known cardiotonic steroids or the substances previously reported to exhibit Na-pump inhibitory activity. Thus, uroxin may be a new type of endogenous regulator for the Na pump.
Tamura
M
M
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146.
Konishi
F
F
Inagami
T
T
D000311:Adrenal Glands / Q000737:chemistry; D000818:Animals; D001463:Barbiturates / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology / Q000652:urine*; D001921:Brain / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008297:Male; D010042:Ouabain / Q000378:metabolism / Q000494:pharmacology; D010446:Peptide Fragments; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D011333:Pro-Opiomelanocortin / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012413:Rubidium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors* / Q000652:urine; D013552:Swine
D001463:Barbiturates; D010446:Peptide Fragments; D010455:Peptides; C078895:joining peptide, pro-opiomelanocortin; D010042:Ouabain; D011333:Pro-Opiomelanocortin; D000254:Sodium-Potassium-Exchanging ATPase; C004469:alloxantin; D012413:Rubidium