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.

Basic information

Original name

HSPA1A conformational mutants reveal a conserved structural unit in Hsp70 proteins

Authors

VANDOVÁ, Veronika (203 Czech Republic), Pavla VAŇKOVÁ, Michal ĎURECH (703 Slovakia), Josef HOUSER (203 Czech Republic, guarantor, belonging to the institution), Daniel KAVAN, Petr MAN, Petr MÜLLER (203 Czech Republic) and Filip TRCKA

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.770

RIV identification code

RIV/00216224:14740/20:00115316

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1016/j.bbagen.2019.129458

UT WoS

000501643100014

Keywords in English

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

Abstract

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.

Links

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

90043, large research infrastructures

Name: CIISB

Citovat

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. 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.

@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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