Biochemical and cytochemical evidence for the polar concentration of periplasmic enzymes in a "minicell" strain of Escherichia coli.
A number of "surface" enzymes of Escherichia coli (i.e., among those selectively released by osmotic shock) all displayed higher specific activities in extracts of minicells than in extracts of typical rod forms; these enzymes included alkaline phosphatase, cyclic phosphodiesterase, acid hexose monophosphatase, 5'-nucleotidase, and ribonuclease I. In addition, alkaline phosphatase, cyclic phosphodiesterase, and acid hexose monophosphatase were cytochemically localized to regions of minicell periplasm that resembled reactive polar enlargements of the periplasm in rod forms. In contrast, a number of "internal" cytoplasmic enzymes (inorganic pyrophosphatase, beta-galactosidase, glutamine synthetase, polynucleotide phosphorylase, and ribonuclease II) showed elevated or similar specific activities in extracts of rod forms versus extracts of minicells. A specific heat-labile inhibitor for 5'-nucleotidase, known to occur in the cytoplasm, also showed no enrichment in minicells. These findings indicate that the "surface" enzymes are segregated in vivo into the terminal minicell buds, possibly because these enzymes are concentrated in the polar enlargements of the periplasm in typical rod forms.
D000469:Alkaline Phosphatase / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology* / Q000201:enzymology* / Q000254:growth & development*; D005696:Galactosidases / Q000378:metabolism*; D005952:Glucose-6-Phosphatase / Q000378:metabolism*; D006651:Histocytochemistry; D006867:Hydrolases / Q000378:metabolism*; D008025:Ligases / Q000378:metabolism*; D008854:Microscopy, Electron; D009708:Nucleotidases / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism*; D011755:Pyrophosphatases / Q000378:metabolism*; D012260:Ribonucleases / Q000302:isolation & purification* / Q000378:metabolism*
D006867:Hydrolases; D012260:Ribonucleases; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D010744:Phosphoric Monoester Hydrolases; D005952:Glucose-6-Phosphatase; D005696:Galactosidases; D011755:Pyrophosphatases; D008025:Ligases