title: The pro- and mature forms of the E. coli K-12 outer membrane phospholipase A are identical.
abstract: The nucleotide sequence of the pldA gene, coding for the outer membrane (OM) phospholipase A of Escherichia coli K-12, and flanking sequences, was determined. Data were obtained from sequences of overlapping deletions which had been generated in vitro from both ends of the gene, using DNase I in the presence of Mn2+ and Bal31 nuclease. The deduced amino acid sequence of the pldA gene product is the first primary sequence of a membrane-bound phospholipase. The complete PldA protein contains 260 amino acids, which include a putative signal sequence, and has a calculated mol. wt. of 29 946 similar to that of the purified protein. Furthermore we found the N terminus of the purified protein to be blocked and the overall amino acid composition to be consistent with the one deduced from the complete pldA gene. Analysis of proteins synthesized in minicells with a pldA coding plasmid in the presence of 8% ethanol did not reveal any new bands on polyacrylamide gels, whereas the control beta-lactamase clearly showed its unprocessed form under the same conditions. These data are consistent with the empirical prediction from the primary sequence, that the PldA protein lacks any signal peptidase 'target' site. We therefore conclude that the PldA protein is exported to the OM without proteolytic removal of the signal peptide.
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004792:Enzyme Precursors / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D010740:Phospholipases / Q000235:genetics*; D010741:Phospholipases A / Q000235:genetics*; D011499:Protein Processing, Post-Translational
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004792:Enzyme Precursors; D010740:Phospholipases; D010741:Phospholipases A