title: Minimum substrate sequence for signal peptidase I of Escherichia coli.
abstract: The minimum substrate sequence recognized by signal peptidase I (SPase I or leader peptidase) was defined by measuring the kinetic parameters for a set of chemically synthesized peptides corresponding to the cleavage site of the precursor maltose binding protein (pro-MBP). The minimum sequence of a substrate hydrolyzed by SPase I at a measurable rate was the pentapeptide Ala-Leu-Ala decreases Lys-Ile. The rates of hydrolysis of this substrate, however, were several hundred-fold lower than those observed for the maturation of MBP in Escherichia coli, suggesting that in addition to these minimal sites involved in recognition, other features of pro-MBP are also needed for the optimal rate of signal peptide cleavage by SPase I. One parameter may be the length of the polypeptide chain. Studies of the synthetic peptides showed that decreasing the length of the polypeptide chain of substrates decreased the substrate efficiency measured as kcat/Km. However, in one case a decrease in the length of a peptide corresponding to -7 to +3 positions of pro-MBP to a nonapeptide (-7 to +2) increased the substrate efficiency by about 900-fold. The nonapeptide is the most efficient substrate for the enzyme in vitro so far reported. It is speculated that better peptide substrates are the ones which are able to adopt folded structures.
authors.affiliation: Division of Molecular Genetics and Microbiology, Burroughs Wellcome & Co., Research Triangle Park, North Carolina 27709.
authors.forename: I K
authors.initials: IK
authors.lastname: Dev
authors.forename: P H
authors.initials: PH
authors.lastname: Ray
authors.forename: P
authors.initials: P
authors.lastname: Novak
mesh_terms: D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D009842:Oligopeptides; D012697:Serine Endopeptidases; D013379:Substrate Specificity
chemical_list: D008565:Membrane Proteins; D009842:Oligopeptides; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase