Disruption of Vi bacteriophage III and localization of its deacetylase activity.
It has been shown that particles of Vi bacteriophage III catalyse deacetylation of O-acetyl pectic (polygalacturonic) acid, a structural analogue of Vi polysaccharide (Vi antigen). Using this substrate, and determining the acetic acid liberated by gas-liquid chromatogrphy, a method for the estimation of Vi phage deacetylase activity has been developed. Purified particles of Vi phage III were exposed to a variety of mildly dissociative reagents and conditions, and then tested for plaque-forming and for deacetylase activity. They have also been inspected under the electron microscope. Osmotic shock, and incubation in the presence of ethylenediamine tetraacetic acid (greater than or equil 0-01 M), or of L-arginine (0-25 M), were found to cause disintegration of the virions into empty head capsids, deoxyribonucleic acid, and base plates still carrying the spikes. The mixtures of viral fragments exhibited an increased deacetylase activity. Using zonal sedimentation and ion exchange chromatography, the phage fragments obtained by treatment with ethylenediaminetetraacetic acid have been fractionated and the base plates isolated. Amongst the viral components, these structures showed the highest specific deacetylase activity. They had the shape of six-pointed stars (about 9-5 nm inner, and 14-5 nm outer diam.) with a central hole or plug (approximately 3 nm), carrying six spikes, roughly cylindrical organelles of approx. 11 X 4 nm, one at each of the points. Of the polypeptides of six sizes (P.1, about 153,000 daltons; P.2, 91,000; P.3, 71,000; P.4 56,500; P.6, 22,000), detected in whole Vi phage III virions by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, only two, P.2 and P.3 were found in the base plates.
D000085:Acetates / Q000096:biosynthesis; D000115:Acetylesterase / Q000302:isolation & purification*; D000596:Amino Acids / Q000494:pharmacology; D001435:Bacteriophages / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D002954:Citrobacter; D004492:Edetic Acid / Q000494:pharmacology; D004755:Enterobacteriaceae; D006026:Glycoside Hydrolases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009995:Osmosis; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D014574:Uronic Acids / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis / Q000302:isolation & purification
D000085:Acetates; D000596:Amino Acids; D010455:Peptides; D011135:Polysaccharides, Bacterial; D014574:Uronic Acids; D014764:Viral Proteins; D004492:Edetic Acid; D000115:Acetylesterase; D006026:Glycoside Hydrolases