Idealization of the hydrophobic segment of the alkaline phosphatase signal peptide

Abstract

Proteins secreted by prokaryotic cells are synthesized as precursors containing an amino-terminal extension sequence or signal peptide¹. Although these signal peptides share little primary sequence homology, recent studies suggest that they function via common pathways during the transport process² and that a common element may reside in their secondary structural characteristics^3–7. We are investigating the role of an idealized hydrophobic sequence with high potential for α-helix formation in the Escherichia coli alkaline phosphatase signal peptide. Here, amino-acid substitutions were made using site-directed mutagenesis to produce a mutant signal sequence containing nine consecutive leucine residues in the hydrophobic core segment. Transport studies with this mutant precursor indicate that mature alkaline phosphatase is correctly targeted to the E. coli periplasm and that processing of the precursor to the mature form of the enzyme is extremely rapid. In contrast, processing is slowed when the mutant signal sequence is lengthened by the insertion of five additional leucine residues and one serine.