Cargando…

Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90

Cyclophilin 40 (Cyp40) comprises an N-terminal cyclophilin domain with peptidyl-prolyl isomerase (PPIase) activity and a C-terminal tetratricopeptide repeat (TPR) domain that binds to the C-terminal–EEVD sequence common to both heat shock protein 70 (Hsp70) and Hsp90. We show in the present study th...

Descripción completa

Detalles Bibliográficos
Autores principales: Blackburn, Elizabeth A., Wear, Martin A., Landré, Vivian, Narayan, Vikram, Ning, Jia, Erman, Burak, Ball, Kathryn L., Walkinshaw, Malcolm D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4721547/
https://www.ncbi.nlm.nih.gov/pubmed/26330616
http://dx.doi.org/10.1042/BSR20150124
_version_ 1782411241970466816
author Blackburn, Elizabeth A.
Wear, Martin A.
Landré, Vivian
Narayan, Vikram
Ning, Jia
Erman, Burak
Ball, Kathryn L.
Walkinshaw, Malcolm D.
author_facet Blackburn, Elizabeth A.
Wear, Martin A.
Landré, Vivian
Narayan, Vikram
Ning, Jia
Erman, Burak
Ball, Kathryn L.
Walkinshaw, Malcolm D.
author_sort Blackburn, Elizabeth A.
collection PubMed
description Cyclophilin 40 (Cyp40) comprises an N-terminal cyclophilin domain with peptidyl-prolyl isomerase (PPIase) activity and a C-terminal tetratricopeptide repeat (TPR) domain that binds to the C-terminal–EEVD sequence common to both heat shock protein 70 (Hsp70) and Hsp90. We show in the present study that binding of peptides containing the MEEVD motif reduces the PPIase activity by ∼30%. CD and fluorescence assays show that the TPR domain is less stable than the cyclophilin domain and is stabilized by peptide binding. Isothermal titration calorimetry (ITC) shows that the affinity for the–MEEVD peptide is temperature sensitive in the physiological temperature range. Results from these biophysical studies fit with the MD simulations of the apo and holo (peptide-bound) structures which show a significant reduction in root mean square (RMS) fluctuation in both TPR and cyclophilin domains when–MEEVD is bound. The MD simulations of the apo-protein also highlight strong anti-correlated motions between residues around the PPIase-active site and a band of residues running across four of the seven helices in the TPR domain. Peptide binding leads to a distortion in the shape of the active site and a significant reduction in these strongly anti-correlated motions, providing an explanation for the allosteric effect of ligand binding and loss of PPIase activity. Together the experimental and MD results suggest that on heat shock, dissociation of Cyp40 from complexes mediated by the TPR domain leads to an increased pool of free Cyp40 capable of acting as an isomerase/chaperone in conditions of cellular stress.
format Online
Article
Text
id pubmed-4721547
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Portland Press Ltd.
record_format MEDLINE/PubMed
spelling pubmed-47215472016-02-02 Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90 Blackburn, Elizabeth A. Wear, Martin A. Landré, Vivian Narayan, Vikram Ning, Jia Erman, Burak Ball, Kathryn L. Walkinshaw, Malcolm D. Biosci Rep Original Papers Cyclophilin 40 (Cyp40) comprises an N-terminal cyclophilin domain with peptidyl-prolyl isomerase (PPIase) activity and a C-terminal tetratricopeptide repeat (TPR) domain that binds to the C-terminal–EEVD sequence common to both heat shock protein 70 (Hsp70) and Hsp90. We show in the present study that binding of peptides containing the MEEVD motif reduces the PPIase activity by ∼30%. CD and fluorescence assays show that the TPR domain is less stable than the cyclophilin domain and is stabilized by peptide binding. Isothermal titration calorimetry (ITC) shows that the affinity for the–MEEVD peptide is temperature sensitive in the physiological temperature range. Results from these biophysical studies fit with the MD simulations of the apo and holo (peptide-bound) structures which show a significant reduction in root mean square (RMS) fluctuation in both TPR and cyclophilin domains when–MEEVD is bound. The MD simulations of the apo-protein also highlight strong anti-correlated motions between residues around the PPIase-active site and a band of residues running across four of the seven helices in the TPR domain. Peptide binding leads to a distortion in the shape of the active site and a significant reduction in these strongly anti-correlated motions, providing an explanation for the allosteric effect of ligand binding and loss of PPIase activity. Together the experimental and MD results suggest that on heat shock, dissociation of Cyp40 from complexes mediated by the TPR domain leads to an increased pool of free Cyp40 capable of acting as an isomerase/chaperone in conditions of cellular stress. Portland Press Ltd. 2015-10-19 /pmc/articles/PMC4721547/ /pubmed/26330616 http://dx.doi.org/10.1042/BSR20150124 Text en © 2015 Authors http://creativecommons.org/licenses/by/3.0/ This is an open access article published by Portland Press Limited and distributed under the Creative Commons Attribution License 3.0 (http://creativecommons.org/licenses/by/3.0/)
spellingShingle Original Papers
Blackburn, Elizabeth A.
Wear, Martin A.
Landré, Vivian
Narayan, Vikram
Ning, Jia
Erman, Burak
Ball, Kathryn L.
Walkinshaw, Malcolm D.
Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90
title Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90
title_full Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90
title_fullStr Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90
title_full_unstemmed Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90
title_short Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90
title_sort cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with hsp90
topic Original Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4721547/
https://www.ncbi.nlm.nih.gov/pubmed/26330616
http://dx.doi.org/10.1042/BSR20150124
work_keys_str_mv AT blackburnelizabetha cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT wearmartina cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT landrevivian cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT narayanvikram cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT ningjia cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT ermanburak cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT ballkathrynl cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90
AT walkinshawmalcolmd cyclophilin40isomeraseactivityisregulatedbyatemperaturedependentallostericinteractionwithhsp90