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

Impakt faktor

Impact factor: 3.770

Kód RIV

RIV/00216224:14740/20:00115316

Organizační jednotka

Středoevropský technologický institut

UT WoS

000501643100014

Klíčová slova anglicky

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

Štítky

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