PURIFICATION OF AN ENDOGENOUS-DIGITALIS-LIKE-FACTOR FROM HUMAN PLASMA FOR STRUCTURAL-ANALYSIS

被引：155

作者：

LUDENS, JH ^{[1
]}

CLARK, MA ^{[1
]}

DUCHARME, DW ^{[1
]}

HARRIS, DW ^{[1
]}

LUTZKE, BS ^{[1
]}

MANDEL, F ^{[1
]}

MATHEWS, WR ^{[1
]}

SUTTER, DM ^{[1
]}

HAMLYN, JM ^{[1
]}

机构：

[1] UNIV MARYLAND,SCH MED,DEPT PHYSIOL,BALTIMORE,MD 21201

来源：

HYPERTENSION | 1991年 / 17卷 / 06期

关键词：

DIGITALIS-LIKE FACTOR; OUABAIN-LIKE COMPOUND; PLASMA; SODIUM PUMP INHIBITOR; SODIUM-POTASSIUM ATPASE;

D O I：

10.1161/01.HYP.17.6.923

中图分类号：

R6 [外科学];

学科分类号：

1002 ; 100210 ;

摘要：

In previous reports, we described the isolation and characterization of an endogenous digitalislike factor (EDLF). In this report, we describe a unique combination of bioassay and large-scale purification methodology that made possible the purification of sufficient quantities of this inhibitor of Na+,K+-ATPase for structural analysis. Using an initial XAD-2 extraction and preparative high-performance liquid chromatography followed by a batch enzyme affinity extraction and two subsequent semipreparative chromatographic steps, 300 1 of human plasma was processed, yielding 31-mu-g (53 nmol) of pure EDLF and representing purification on a dry weight basis in excess of 0.6 billionfold. Four divergent pieces of evidence, including chromatographic, mass spectrometric, immunoreactive, and binding characteristics, suggested that the EDLF purified in the present study was either ouabain or an isomer of ouabain. This material may represent a plasma-borne, naturally occurring, selective, high-affinity ligand for the digitalis binding site that may play a significant role in the modulation of the sodium pump and thereby cellular electrolyte homeostasis in humans.

引用

页码：923 / 929

页数：7

共 28 条

[1] EFFECTS OF AN ENDOGENOUS OUABAIN-LIKE COMPOUND ON HEART AND AORTA [J].