An Escherichia coli twin-arginine signal peptide switches between helical and unstructured conformations depending on the hydrophobicity of the environment

Miguel San Miguel, Rachel Marrington, P. Mark Rodger, Alison Rodger, Colin Robinson*

*Corresponding author for this work

Research output: Contribution to journalArticle

32 Citations (Scopus)

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% α-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 non-functional 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.

Original languageEnglish
Pages (from-to)3345-3352
Number of pages8
JournalEuropean Journal of Biochemistry
Volume270
Issue number16
DOIs
Publication statusPublished - Aug 2003
Externally publishedYes

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Keywords

  • Protein transport
  • Signal peptide
  • SufI
  • Tat system
  • Twin-arginine translocation

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