title: An Escherichia coli twin-arginine signal peptide switches between helical and unstructured conformations depending on the hydrophobicity of the environment.
abstract: The Tat system catalyzes the transport of folded globular proteins across the bacterial plasma membrane and the chloroplast thylakoid. It recognizes cleavable signal peptides containing a critical twin-arginine motif but little is known of the overall structure of these peptides. In this report, we have analyzed the secondary structure of the SufI signal peptide, together with those of two nonfunctional variants in which the region around the twin-arginine, RRQFI, is replaced by KKQFI or RRQAA. Circular dichroism studies show that the SufI peptide exists as an unstructured peptide in aqueous solvent with essentially no stable secondary structure. In membrane-mimetic environments such as SDS micelles or water/trifluoroethanol, however, the peptide adopts a structure containing up to about 40% alpha-helical content. Secondary structure predictions and molecular modelling programs strongly suggest that the helical region begins at, or close to, the twin-arginine motif. Studies on the thermal stability of the helix demonstrate a sharp transition between the unstructured and helical states, suggesting that the peptide exists in one of two distinct states. The two nonfunctional peptides exhibit almost identical spectra and properties to the wild-type SufI peptide, indicating that it is the arginine sidechains, and not their contribution to the helical structure, that are critical in this class of peptide.
authors.affiliation: Department of Biological Sciences and Department of Chemistry, University of Warwick, Coventry, UK.
authors.forename: Miguel
authors.initials: M
authors.lastname: San Miguel
authors.forename: Rachel
authors.initials: R
authors.lastname: Marrington
authors.forename: P Mark
authors.initials: PM
authors.lastname: Rodger
authors.forename: Alison
authors.initials: A
authors.lastname: Rodger
authors.forename: Colin
authors.initials: C
authors.lastname: Robinson
mesh_terms: D001120:Arginine / Q000378:metabolism*; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000502:physiology*; D017433:Protein Structure, Secondary
chemical_list: D010455:Peptides; D021382:Protein Sorting Signals; D001120:Arginine