Overlapping transport and chaperone-binding functions within a bacterial twin-arginine signal peptide.

Molecular Microbiology
Sabine GrahlFrank Sargent

Abstract

The twin-arginine translocation (Tat) pathway is a protein targeting system present in many prokaryotes. The physiological role of the Tat pathway is the transmembrane translocation of fully-folded proteins, which are targeted by N-terminal signal peptides bearing conserved SRRxFLK 'twin-arginine' amino acid motifs. In Escherichia coli the majority of Tat targeted proteins bind redox cofactors and it is important that only mature, cofactor-loaded precursors are presented for export. Cellular processes have been unearthed that sequence these events, for example the signal peptide of the periplasmic nitrate reductase (NapA) is bound by a cytoplasmic chaperone (NapD) that is thought to regulate assembly and export of the enzyme. In this work, genetic, biophysical and structural approaches were taken to dissect the interaction between NapD and the NapA signal peptide. A NapD binding epitope was identified towards the N-terminus of the signal peptide, which overlapped significantly with the twin-arginine targeting motif. NMR spectroscopy revealed that the signal peptide adopted a α-helical conformation when bound by NapD, and substitution of single residues within the NapA signal peptide was sufficient to disrupt the interaction. Th...Continue Reading

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Citations

Oct 16, 2015·FEMS Microbiology Reviews·Silke Leimkühler, Chantal Iobbi-Nivol
Aug 22, 2013·Molecular Microbiology·Martyn J JamesFrank Sargent
Dec 4, 2012·Biochimica Et Biophysica Acta·Chantal Iobbi-Nivol, Silke Leimkühler
Jul 31, 2012·Biochimica Et Biophysica Acta·Jörg Simon, Martin G Klotz
Oct 6, 2015·EcoSal Plus·Axel Magalon, Ralf R Mendel
May 25, 2021·Frontiers in Molecular Biosciences·Sylvain Durand, Maude Guillier

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Methods Mentioned

BETA
nuclear magnetic resonance
two-hybrid
isothermal titration calorimetry
NMR
PCR
two hybrid
two

Software Mentioned

Origin
TALOS
YASARA
NMRPipe
YASARA2
NapD
TALOS +
SPARKY

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