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
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
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