K99 surface antigen of Escherichia coli: purification and partial characterization.
K99, a presumed colonizing factor of enterotoxigenic Escherichia coli of calf origin, has been purified. K99 was removed from K99+ bacteria by salt extraction and subsequently purified by ammonium sulfate precipitation and column chromatography on diethylaminoethyl-Sephadex. The purified material was homogenous in size, having an s20,w of 13 to 15 S. It was composed of two subunits: a major component with a molecular weight of 22,500 and a minor component of 29,500. When observed in the electron microscope, K99 appeared to be rod-shaped, with a strong tendency for self-aggregation. At concentrations where aggregation was minimized, individual rods were observed with diameters of 8.4 nm and mean lengths of 130 nm. Based on the subunit structure, exterior location, and rod-like shape of K99, it is concluded that K99 is a pilus or pilus-like structure. Chemically, K99 is composed primarily of protein and has an isoelectric point of greater than 10. Purified K99 did not hemagglutinate guinea pig erythrocytes.
D000645:Ammonium Sulfate / Q000494:pharmacology; D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D002417:Cattle; D002458:Cell Fractionation; D002462:Cell Membrane / Q000276:immunology*; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004926:Escherichia coli / Q000276:immunology* / Q000648:ultrastructure; D006384:Hemagglutination; D005779:Immunodiffusion; D007525:Isoelectric Focusing; D008970:Molecular Weight; D014461:Ultracentrifugation
D000942:Antigens, Bacterial; D000645:Ammonium Sulfate