Isolation and purification of the envelope proteins of Newcastle disease virus.
A procedure has been developed for the isolation of Newcastle disease virus (NDV) envelope proteins. The two surface glycoproteins and the non-glycosylated membrane protein were solubilized with 2% Triton X-100 and 1 m KCl. Removal of the KCl by dialysis yielded by precipitation a pure preparation of the non-glycosylated membrane protein, which is insoluble in solutions of low ionic strength. The soluble fraction consisting of the two glycoproteins possessed full neuraminidase and hemagglutinating activities. The two glycoproteins could be separated by rate zonal sedimentation in a sucrose gradient containing 1% Triton X-100 and 1 m KCl. Under these conditions, the sedimentation coefficient of the larger glycoprotein, virus protein 1, was 9.3s, and that of the smaller, virus protein 2, was 6.1s. Both hemagglutinating and neuraminidase activities were associated with virus protein 1; virus protein 2 had neither activity. The results suggest that both activities reside on a single NDV glycoprotein. Similar results were obtained previously with another paramyxovirus, simian virus 5. These findings suggest that the association of hemagglutinating and neuraminidase activities with one glycoprotein is a general property of the paramyxovirus group.
D000596:Amino Acids; D000818:Animals; D002247:Carbon Isotopes; D002417:Cattle; D002460:Cell Line; D002501:Centrifugation, Zonal; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis / Q000302:isolation & purification / Q000494:pharmacology; D006386:Hemagglutination Tests; D007668:Kidney; D007930:Leucine; D008722:Methods; D009439:Neuraminidase / Q000378:metabolism; D009522:Newcastle disease virus / Q000032:analysis* / Q000201:enzymology / Q000254:growth & development / Q000276:immunology; D011189:Potassium Chloride; D013501:Surface-Active Agents; D014316:Tritium; D014764:Viral Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology
D000596:Amino Acids; D002247:Carbon Isotopes; D006023:Glycoproteins; D013501:Surface-Active Agents; D014764:Viral Proteins; D014316:Tritium; D011189:Potassium Chloride; D009439:Neuraminidase; D007930:Leucine