Comparative study on fibers isolated from four R-type pyocins, phage-tail-like bacteriocins of Pseudomonas aeruginosa.
Five R-type pyocins have been reported which are almost identical with one another in their morphology and subunit composition, though distinct in receptor-binding specificity. We isolated fibers from pyocin R2, R3, and R4 by essentially the same procedure as used in our previous isolation of pyocin R1 fiber with unimpaired receptor-binding ability. All the isolated fibers including R1 fiber were indistinguishable from one another, in terms of electron microscopic observation and subunit composition analysis by SDS gel electrophoresis. They consisted mainly of Subunit No. 2 (Mw 71,000) and No. 9 (31,000) proteins. Although Subunit No. 9 protein in every fiber was susceptible to trypsin and afforded a fragment with the same molecular weight (about 19,000) detectable in the SDS gel, Subunit No. 2 protein was cleaved with trypsin only after the fiber had been treated with an organomercurial, 4-(p-sulfophenylazo)-2-mercuriphenol. The cleavage of Subunit No. 2 protein proceeded to give several fragments with molecular weights ranging from 64,000 to 34,000, and the fragmentation patterns were electrophoretically distinct at least among R1 fiber, R3 fiber, and others (R2 and R4 fibers). The results indicate that Subunit No. 2 proteins of these fibers are different from one another in the structure surrounding trypsin-susceptible peptide bonds. Immunological investigations with anti-R1 fiber antibodies provided some additional information on the difference among R-type pyocins at the fiber level.
D000596:Amino Acids / Q000032:analysis; D000936:Antigen-Antibody Complex; D001430:Bacteriocins / Q000302:isolation & purification*; D007106:Immune Sera; D005779:Immunodiffusion; D008854:Microscopy, Electron; D010446:Peptide Fragments / Q000032:analysis; D011550:Pseudomonas aeruginosa / Q000032:analysis*; D011709:Pyocins / Q000302:isolation & purification*; D014357:Trypsin
D000596:Amino Acids; D000936:Antigen-Antibody Complex; D001430:Bacteriocins; D007106:Immune Sera; D010446:Peptide Fragments; D011709:Pyocins; D014357:Trypsin