HSPA1A conformational mutants reveal a conserved structural
unit in Hsp70 proteins

J 2020

HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins

VANDOVÁ, Veronika; Pavla VAŇKOVÁ; Michal ĎURECH; Josef HOUSER; Daniel KAVAN et. al.

Základní údaje

Originální název

HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins

Autoři

VANDOVÁ, Veronika (203 Česká republika); Pavla VAŇKOVÁ; Michal ĎURECH (703 Slovensko); Josef HOUSER (203 Česká republika, garant, domácí); Daniel KAVAN; Petr MAN; Petr MÜLLER (203 Česká republika) a Filip TRCKA

Vydání

Biochimica et Biophysica Acta - General Subjects, Amsterdam, Elsevier, 2020, 0304-4165

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.770

Kód RIV

RIV/00216224:14740/20:00115316

Organizační jednotka

Středoevropský technologický institut

DOI

https://doi.org/10.1016/j.bbagen.2019.129458

UT WoS

000501643100014

EID Scopus

2-s2.0-85074488024

Klíčová slova anglicky

Allostery; Molecular chaperones; Heat-shock protein 70; Folding; Mutation

Štítky

CF BIC, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 27. 10. 2024 15:08, Ing. Martina Blahová

Anotace

V originále

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.

Návaznosti

90043, velká výzkumná infrastruktura

Název: CIISB

Citovat

VANDOVÁ, Veronika; Pavla VAŇKOVÁ; Michal ĎURECH; Josef HOUSER; Daniel KAVAN; Petr MAN; Petr MÜLLER a Filip TRCKA. HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins. Biochimica et Biophysica Acta - General Subjects. Amsterdam: Elsevier, 2020, roč. 1864, č. 1, s. 1-16. ISSN 0304-4165. Dostupné z: https://doi.org/10.1016/j.bbagen.2019.129458.

@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 = {https://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 a Filip TRCKA. HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins. \textit{Biochimica et Biophysica Acta - General Subjects}. Amsterdam: Elsevier, 2020, roč.~1864, č.~1, s.~1-16. ISSN~0304-4165. Dostupné z: https://doi.org/10.1016/j.bbagen.2019.129458.

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