Studies on the modification and processing of prolipoprotein in Escherichia coli. Effects of structural alterations in prolipoprotein on its maturation in wild type and lpp mutants.
We have previously shown that an Escherichia coli mutant ( mlpA allele) containing a structurally altered murein prolipoprotein due to substitution of Gly14 by Asp14 , is globomycin resistant. In addition, the mutant prolipoprotein is not modified with glyceride and consequently remains uncleaved. Spontaneous revertants possessing a mature lipoprotein of apparent normal structure can be isolated by EDTA selection. Three revertants were chosen in the present study which included the analysis of kinetics of lipoprotein maturation and the determination of globomycin sensitivity. These pseudorevertants in the lpp gene which could be recognized by the anomalous prolipoprotein mobility in sodium dodecyl sulfate gels, exhibited altered globomycin sensitivity in vivo. Our results indicate that alterations in prolipoprotein structure affect the kinetics of prolipoprotein modification and processing reactions, both in vivo and in vitro. Pulse-chase experiments revealed the transient existence of unmodified prolipoprotein and modified prolipoprotein as biosynthetic intermediates of mature lipoprotein. The rate of prolipoprotein modification appeared to be slightly faster than that of processing in the wild type cell. In contrast, modification of prolipoprotein was rate limiting in a pseudorevertant strain 14R21 , and the processing of 14R21 modified prolipoprotein appeared to proceed more rapidly than that of wild type prolipoprotein, both in vitro and in vivo.
D000900:Anti-Bacterial Agents / Q000633:toxicity; D001425:Bacterial Outer Membrane Proteins; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D005838:Genotype; D007700:Kinetics; D008074:Lipoproteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010455:Peptides / Q000633:toxicity; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity
D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D008074:Lipoproteins; C099932:Lpp protein, E coli; D010455:Peptides; D011498:Protein Precursors; C017339:globomycin