Studies on biologically Active Substances from Rabbit Skin Tissues infected with Vaccinia virus.-Isolation and Properties of Inhibitor to Gastric Acid Secretion

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Abstract
Methods are described for the isolation of biologically active substances from rabbit skin tissues infected with vaccinia virus. The inhibitory activity on tetragastrin-stimulated gastric acid secretion found in the alkaline aqueous extract of the infected tissues boiled in acetone was concentrated by precipitation with (NH4)2SO4 and acetone. Further purification of the active material was carried out by chromatography over DEAE-cellulose and gel filtration over Sephadex G-75. The yield of the purified protein, (I) was 2.25 mg from 1.0 kg of infected tissues and showed 77.2 .+-. 8.4% (n = 5) of inhibitory activity when doses of 5.0 mg/kg were administered i.v. into rats. This protein was also a potent inhibitor of gastric acid, gastric juice, pepsin secretion and ulcers induced by pylorus ligation. MW of I was 69,000 by sodium dodecyl sulfate gel electrophoresis and gel filtration through Sephadex G-75. Amino acid analysis demonstrated 18 different amino acids with high content of glutamic acid and proline. Another biologically active material (II) was extracted with boiled aqueous phenol and fractionated with Dowex 50W chromatography. This material contained uracil, xanthine, several amino acid-related substances and nucleoside-related substance. Ornithine, ethanolamine and II inhibited heat-induced hemolysis of rabbit erythrocytes, and II was also a potent inhibitor of chemotaxis of rabbit polymorphonuclears in vitro.

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