Cargando…
Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase
Protein domains often recognize short linear protein motifs composed of a core conserved consensus sequence surrounded by less critical, modulatory positions. PTEN, a lipid phosphatase involved in phosphatidylinositol 3-kinase (PI3K) pathway, contains such a short motif located at the extreme C-term...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7774954/ https://www.ncbi.nlm.nih.gov/pubmed/33382810 http://dx.doi.org/10.1371/journal.pone.0244613 |
_version_ | 1783630370081603584 |
---|---|
author | Jané, Pau Gógl, Gergő Kostmann, Camille Bich, Goran Girault, Virginie Caillet-Saguy, Célia Eberling, Pascal Vincentelli, Renaud Wolff, Nicolas Travé, Gilles Nominé, Yves |
author_facet | Jané, Pau Gógl, Gergő Kostmann, Camille Bich, Goran Girault, Virginie Caillet-Saguy, Célia Eberling, Pascal Vincentelli, Renaud Wolff, Nicolas Travé, Gilles Nominé, Yves |
author_sort | Jané, Pau |
collection | PubMed |
description | Protein domains often recognize short linear protein motifs composed of a core conserved consensus sequence surrounded by less critical, modulatory positions. PTEN, a lipid phosphatase involved in phosphatidylinositol 3-kinase (PI3K) pathway, contains such a short motif located at the extreme C-terminus capable to recognize PDZ domains. It has been shown that the acetylation of this motif could modulate the interaction with several PDZ domains. Here we used an accurate experimental approach combining high-throughput holdup chromatographic assay and competitive fluorescence polarization technique to measure quantitative binding affinity profiles of the PDZ domain-binding motif (PBM) of PTEN. We substantially extended the previous knowledge towards the 266 known human PDZ domains, generating the full PDZome-binding profile of the PTEN PBM. We confirmed that inclusion of N-terminal flanking residues, acetylation or mutation of a lysine at a modulatory position significantly altered the PDZome-binding profile. A numerical specificity index is also introduced as an attempt to quantify the specificity of a given PBM over the complete PDZome. Our results highlight the impact of modulatory residues and post-translational modifications on PBM interactomes and their specificity. |
format | Online Article Text |
id | pubmed-7774954 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-77749542021-01-11 Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase Jané, Pau Gógl, Gergő Kostmann, Camille Bich, Goran Girault, Virginie Caillet-Saguy, Célia Eberling, Pascal Vincentelli, Renaud Wolff, Nicolas Travé, Gilles Nominé, Yves PLoS One Research Article Protein domains often recognize short linear protein motifs composed of a core conserved consensus sequence surrounded by less critical, modulatory positions. PTEN, a lipid phosphatase involved in phosphatidylinositol 3-kinase (PI3K) pathway, contains such a short motif located at the extreme C-terminus capable to recognize PDZ domains. It has been shown that the acetylation of this motif could modulate the interaction with several PDZ domains. Here we used an accurate experimental approach combining high-throughput holdup chromatographic assay and competitive fluorescence polarization technique to measure quantitative binding affinity profiles of the PDZ domain-binding motif (PBM) of PTEN. We substantially extended the previous knowledge towards the 266 known human PDZ domains, generating the full PDZome-binding profile of the PTEN PBM. We confirmed that inclusion of N-terminal flanking residues, acetylation or mutation of a lysine at a modulatory position significantly altered the PDZome-binding profile. A numerical specificity index is also introduced as an attempt to quantify the specificity of a given PBM over the complete PDZome. Our results highlight the impact of modulatory residues and post-translational modifications on PBM interactomes and their specificity. Public Library of Science 2020-12-31 /pmc/articles/PMC7774954/ /pubmed/33382810 http://dx.doi.org/10.1371/journal.pone.0244613 Text en © 2020 Jané et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Jané, Pau Gógl, Gergő Kostmann, Camille Bich, Goran Girault, Virginie Caillet-Saguy, Célia Eberling, Pascal Vincentelli, Renaud Wolff, Nicolas Travé, Gilles Nominé, Yves Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase |
title | Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase |
title_full | Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase |
title_fullStr | Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase |
title_full_unstemmed | Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase |
title_short | Interactomic affinity profiling by holdup assay: Acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase |
title_sort | interactomic affinity profiling by holdup assay: acetylation and distal residues impact the pdzome-binding specificity of pten phosphatase |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7774954/ https://www.ncbi.nlm.nih.gov/pubmed/33382810 http://dx.doi.org/10.1371/journal.pone.0244613 |
work_keys_str_mv | AT janepau interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT goglgergo interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT kostmanncamille interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT bichgoran interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT giraultvirginie interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT cailletsaguycelia interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT eberlingpascal interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT vincentellirenaud interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT wolffnicolas interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT travegilles interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase AT nomineyves interactomicaffinityprofilingbyholdupassayacetylationanddistalresiduesimpactthepdzomebindingspecificityofptenphosphatase |