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The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition

BACKGROUND: The majority of peroxisomal matrix proteins destined for translocation into the peroxisomal lumen are recognised via a C-terminal Peroxisomal Target Signal type 1 by the cycling receptor Pex5p. The only structure to date of Pex5p in complex with a cargo protein is that of the C-terminal...

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Autores principales: Williams, Chris P, Schueller, Nicole, Thompson, Colin A, van den Berg, Marlene, Van Haren, Simon D, Erdmann, Ralf, Bond, Charles S, Distel, Ben, Schliebs, Wolfgang, Wilmanns, Matthias, Stanley, Will A
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3060121/
https://www.ncbi.nlm.nih.gov/pubmed/21375735
http://dx.doi.org/10.1186/1471-2091-12-12
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author Williams, Chris P
Schueller, Nicole
Thompson, Colin A
van den Berg, Marlene
Van Haren, Simon D
Erdmann, Ralf
Bond, Charles S
Distel, Ben
Schliebs, Wolfgang
Wilmanns, Matthias
Stanley, Will A
author_facet Williams, Chris P
Schueller, Nicole
Thompson, Colin A
van den Berg, Marlene
Van Haren, Simon D
Erdmann, Ralf
Bond, Charles S
Distel, Ben
Schliebs, Wolfgang
Wilmanns, Matthias
Stanley, Will A
author_sort Williams, Chris P
collection PubMed
description BACKGROUND: The majority of peroxisomal matrix proteins destined for translocation into the peroxisomal lumen are recognised via a C-terminal Peroxisomal Target Signal type 1 by the cycling receptor Pex5p. The only structure to date of Pex5p in complex with a cargo protein is that of the C-terminal cargo-binding domain of the receptor with sterol carrier protein 2, a small, model peroxisomal protein. In this study, we have tested the contribution of a second, ancillary receptor-cargo binding site, which was found in addition to the characterised Peroxisomal Target Signal type 1. RESULTS: To investigate the function of this secondary interface we have mutated two key residues from the ancillary binding site and analyzed the level of binding first by a yeast-two-hybrid assay, followed by quantitative measurement of the binding affinity and kinetics of purified protein components and finally, by in vivo measurements, to determine translocation capability. While a moderate but significant reduction of the interaction was found in binding assays, we were not able to measure any significant defects in vivo. CONCLUSIONS: Our data therefore suggest that at least in the case of sterol carrier protein 2 the contribution of the second binding site is not essential for peroxisomal import. At this stage, however, we cannot rule out that other cargo proteins may require this ancillary binding site.
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spelling pubmed-30601212011-03-18 The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition Williams, Chris P Schueller, Nicole Thompson, Colin A van den Berg, Marlene Van Haren, Simon D Erdmann, Ralf Bond, Charles S Distel, Ben Schliebs, Wolfgang Wilmanns, Matthias Stanley, Will A BMC Biochem Research Article BACKGROUND: The majority of peroxisomal matrix proteins destined for translocation into the peroxisomal lumen are recognised via a C-terminal Peroxisomal Target Signal type 1 by the cycling receptor Pex5p. The only structure to date of Pex5p in complex with a cargo protein is that of the C-terminal cargo-binding domain of the receptor with sterol carrier protein 2, a small, model peroxisomal protein. In this study, we have tested the contribution of a second, ancillary receptor-cargo binding site, which was found in addition to the characterised Peroxisomal Target Signal type 1. RESULTS: To investigate the function of this secondary interface we have mutated two key residues from the ancillary binding site and analyzed the level of binding first by a yeast-two-hybrid assay, followed by quantitative measurement of the binding affinity and kinetics of purified protein components and finally, by in vivo measurements, to determine translocation capability. While a moderate but significant reduction of the interaction was found in binding assays, we were not able to measure any significant defects in vivo. CONCLUSIONS: Our data therefore suggest that at least in the case of sterol carrier protein 2 the contribution of the second binding site is not essential for peroxisomal import. At this stage, however, we cannot rule out that other cargo proteins may require this ancillary binding site. BioMed Central 2011-03-04 /pmc/articles/PMC3060121/ /pubmed/21375735 http://dx.doi.org/10.1186/1471-2091-12-12 Text en Copyright © 2011 Williams et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Williams, Chris P
Schueller, Nicole
Thompson, Colin A
van den Berg, Marlene
Van Haren, Simon D
Erdmann, Ralf
Bond, Charles S
Distel, Ben
Schliebs, Wolfgang
Wilmanns, Matthias
Stanley, Will A
The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
title The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
title_full The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
title_fullStr The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
title_full_unstemmed The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
title_short The Peroxisomal Targeting Signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
title_sort peroxisomal targeting signal 1 in sterol carrier protein 2 is autonomous and essential for receptor recognition
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3060121/
https://www.ncbi.nlm.nih.gov/pubmed/21375735
http://dx.doi.org/10.1186/1471-2091-12-12
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