Stoichiometry for binding and transport by the twin arginine translocation system.

The Journal of Cell Biology
Jose M Celedon, Kenneth Cline

Abstract

Twin arginine translocation (Tat) systems transport large folded proteins across sealed membranes. Tat systems accomplish this feat with three membrane components organized in two complexes. In thylakoid membranes, cpTatC and Hcf106 comprise a large receptor complex containing an estimated eight cpTatC-Hcf106 pairs. Protein transport occurs when Tha4 joins the receptor complex as an oligomer of uncertain size that is thought to form the protein-conducting structure. Here, binding analyses with intact membranes or purified complexes indicate that each receptor complex could bind eight precursor proteins. Kinetic analysis of translocation showed that each precursor-bound site was independently functional for transport, and, with sufficient Tha4, all sites were concurrently active for transport. Tha4 titration determined that ∼26 Tha4 protomers were required for transport of each OE17 (oxygen-evolving complex subunit of 17 kD) precursor protein. Our results suggest that, when fully saturated with precursor proteins and Tha4, the Tat translocase is an ∼2.2-megadalton complex that can individually transport eight precursor proteins or cooperatively transport multimeric precursors.

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Citations

Dec 20, 2012·Nature Communications·Julia FröbelMatthias Müller
Sep 5, 2013·Proceedings of the National Academy of Sciences of the United States of America·Felicity AlcockBen C Berks
Mar 9, 2013·Proceedings of the National Academy of Sciences of the United States of America·Fernanda RodriguezJason R Schnell
Apr 9, 2014·The Journal of Cell Biology·Cassie AldridgeKenneth Cline
Feb 6, 2013·Biochimica Et Biophysica Acta·René Steffen HauerRalf Bernd Klösgen
Mar 4, 2014·Biochimica Et Biophysica Acta·Roshani PatelColin Robinson
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Oct 21, 2015·The Journal of Biological Chemistry·Ekaterina EimerMatthias Müller
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Mar 17, 2021·Proceedings of the National Academy of Sciences of the United States of America·Anthony H Asher, Steven M Theg
Jun 20, 2019·EcoSal Plus·Kelly M FrainColin Robinson

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Methods Mentioned

BETA
cross-linking studies
affinity purification
immunoprecipitation
electrophoresis
cross-linking study
PCR
protein chase
saturation assay

Software Mentioned

GNU
Prism
GraphPad
LabPlot
Quantity One

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