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@article{1621217,
author = {Vandová, Veronika and Vaňková, Pavla and Ďurech, Michal and Houser, Josef and Kavan, Daniel and Man, Petr and Müller, Petr and Trcka, Filip},
article_location = {Amsterdam},
article_number = {1},
doi = {http://dx.doi.org/10.1016/j.bbagen.2019.129458},
keywords = {Allostery; Molecular chaperones; Heat-shock protein 70; Folding; Mutation},
language = {eng},
issn = {0304-4165},
journal = {Biochimica et Biophysica Acta - General Subjects},
title = {HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins},
url = {https://doi.org/10.1016/j.bbagen.2019.129458},
volume = {1864},
year = {2020}
}
TY - JOUR
ID - 1621217
AU - Vandová, Veronika - Vaňková, Pavla - Ďurech, Michal - Houser, Josef - Kavan, Daniel - Man, Petr - Müller, Petr - Trcka, Filip
PY - 2020
TI - HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins
JF - Biochimica et Biophysica Acta - General Subjects
VL - 1864
IS - 1
SP - 1-16
EP - 1-16
PB - Elsevier
SN - 03044165
KW - Allostery
KW - Molecular chaperones
KW - Heat-shock protein 70
KW - Folding
KW - Mutation
UR - https://doi.org/10.1016/j.bbagen.2019.129458
L2 - https://doi.org/10.1016/j.bbagen.2019.129458
N2 - Background: The Hsp70 proteins maintain proteome integrity through the capacity of their nucleotide- and substrate-binding domains (NBD and SBD) to allosterically regulate substrate affinity in a nucleotide-dependent manner. Crystallographic studies showed that Hsp70 allostery relies on formation of contacts between ATP-bound NBD and an interdomain linker, accompanied by SBD subdomains docking onto distinct sites of the NBD leading to substrate release. However, the mechanics of ATP-induced SBD subdomains detachment is largely unknown. Methods: Here, we investigated the structural and allosteric properties of human HSPA1A using hydrogen/deuterium exchange mass spectrometry, ATPase assays, surface plasmon resonance and fluorescence polarization-based substrate binding assays. Results: Analysis of HSPA1A proteins bearing mutations at the interface of SBD subdomains close to the interdomain linker (amino acids L399, L510, 1515, and D529) revealed that this region forms a folding unit stabilizing the structure of both SBD subdomains in the nucleotide-free state. The introduced mutations modulate HSPA1A allostery as they localize to the NBD-SBD interfaces in the ATP-bound protein. Conclusions: These findings show that residues forming the hydrophobic structural unit stabilizing the SBD structure are relocated during ATP-activated detachment of the SBD subdomains to different NBD-SBD docking interfaces enabling HSPA1A allostery. General significance: Mutation-induced perturbations tuned HSPA1A sensitivity to peptide/protein substrates and to Hsp40 in a way that is common for other Hsp70 proteins. Our results provide an insight into structural rearrangements in the SBD of Hsp70 proteins and highlight HSPA1A-specific allostery features, which is a prerequisite for selective targeting in Hsp-related pathologies.
ER -
VANDOVÁ, Veronika, Pavla VAŇKOVÁ, Michal ĎURECH, Josef HOUSER, Daniel KAVAN, Petr MAN, Petr MÜLLER and Filip TRCKA. HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins. \textit{Biochimica et Biophysica Acta - General Subjects}. Amsterdam: Elsevier, 2020, vol.~1864, No~1, p.~1-16. ISSN~0304-4165. Available from: https://dx.doi.org/10.1016/j.bbagen.2019.129458.
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