Leucocytosis-promoting factor of Bordetella pertussis. I. Purification and characterization.
The leucocytosis-promoting factor was purified from the supernatant fluid of spent cultures of Bordetella pertussis on solid medium. After precipitation at 67% saturation of ammonium sulfate, the leucocytosis-promoting factor was extracted with a 1.0 m NaCl solution. Purification was accomplished by starch block electrophoresis and sucrose density gradient centrifugation. The purified preparation contained a high leucocytosis-promoting activity, and as small an amount as 0.04 mug of protein induced leucocytosis in mice. About 520-fold purification was attained, with a re-recovery of about 25% on an activity basis. The leucocytosis-promoting factor was composed solely of filamentous molecules of about 2 by 40 nm in size, with a sedimentation coefficient of approximately 5.5S and a molecular weight of 108,000. It was insoluble in water but partially soluble in 1.0 m NaCl solution, and consisted mainly of protein, with some carbohydrate, lipid, and phosphorus.
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D001886:Bordetella pertussis / Q000276:immunology*; D002482:Cellulose; D002499:Centrifugation, Density Gradient; D004269:DNA, Bacterial / Q000032:analysis; D004586:Electrophoresis; D004592:Electrophoresis, Starch Gel; D007122:Immunoelectrophoresis; D007964:Leukocytosis / Q000276:immunology*; D051379:Mice; D008854:Microscopy, Electron; D008970:Molecular Weight; D009584:Nitrogen / Q000032:analysis; D012329:RNA, Bacterial / Q000032:analysis; D014461:Ultracentrifugation
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D002482:Cellulose; D009584:Nitrogen