title: A conservative amino acid substitution, arginine for lysine, abolishes export of a hybrid protein in Escherichia coli. Implications for the mechanism of protein secretion.
abstract: A hybrid protein that comprises the beta-lactamase signal peptide fused precisely to chicken muscle triosephosphate isomerase is not secreted into the periplasm of Escherichia coli. The protein can be secreted, however, if an arginine residue at position 3 of the isomerase is replaced by either a serine or a proline residue. In contrast, replacement of a neighboring lysine residue has no effect on secretion of the protein. Furthermore, if the arginine is removed from position 3 to generate a secreted protein, but is then reintroduced in place of the neighboring lysine, the blockade to secretion is re-established. The singular effect of the arginine residue on secretion does not result from the role this residue plays in the formation or stabilization of the native isomerase structure: mutational alterations remote from the N terminus of the isomerase that prevent the proper folding of the protein do not relieve the block to secretion. The finding that an arginine residue prevents secretion while a lysine residue does not, suggests that basic residues near the mature N terminus of a secreted protein must be deprotonated if orderly export is to occur. This implies that the signal peptide along with the N-terminal portion of the mature protein partitions directly into the lipid bilayer in the course of the secretory process.
authors.affiliation: Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138.
authors.forename: R G
authors.initials: RG
authors.lastname: Summers
authors.forename: C R
authors.initials: CR
authors.lastname: Harris
authors.forename: J R
authors.initials: JR
authors.lastname: Knowles
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D002238:Carbohydrate Epimerases / Q000235:genetics*; D002645:Chickens; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D008239:Lysine; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D009154:Mutation; D010957:Plasmids; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014305:Triose-Phosphate Isomerase / Q000096:biosynthesis / Q000235:genetics*; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D001120:Arginine; D001618:beta-Lactamases; D002238:Carbohydrate Epimerases; D014305:Triose-Phosphate Isomerase; D008239:Lysine