title: Effect of amino acid substitutions at the signal peptide cleavage site of the Escherichia coli major outer membrane lipoprotein.
abstract: The requirement for the glycine residue at the COOH terminus of the signal peptide of the precursor of the major Escherichia coli outer membrane lipoprotein was examined. Using oligonucleotide-directed site-specific mutagenesis, this residue was replaced by residues of increasing side chain size. Substitution by serine had no effect on the modification or processing of the prolipoprotein. Substitution by valine or leucine resulted in the accumulation of the unmodified precursor, whereas threonine substitution resulted in slow lipid modification and no detectable processing of the lipid modified precursor. The results indicate that serine is the upper limit on size for the residue at the cleavage site. Larger residues at this position prevent the action of both the glyceride transferase and signal peptidase II enzymes, indicating that the cleavage site residue plays a role in events prior to proteolytic cleavage. The upper limit on size of the cleavage site residue is similar to that found for exported proteins cleaved by signal peptidase I, as well as eucaryotic exported proteins. The possibility that the cleavage site residue may have a role other than active site recognition by the signal peptidase is discussed.
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D008074:Lipoproteins / Q000378:metabolism*; D008565:Membrane Proteins; D011487:Protein Conformation; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000378:metabolism*; D012697:Serine Endopeptidases
chemical_list: D001425:Bacterial Outer Membrane Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D011498:Protein Precursors; D021382:Protein Sorting Signals; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase