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Loss of stability and unfolding cooperativity in hPGK1 upon gradual structural perturbation of its N-terminal domain hydrophobic core
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SYSNO ASEP 0564237 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Loss of stability and unfolding cooperativity in hPGK1 upon gradual structural perturbation of its N-terminal domain hydrophobic core Tvůrce(i) Luis Pacheco-Garcia, J. (ES)
Loginov, Dmitry Sergej (MBU-M) RID
Naganathan, A. N. (IN)
Vaňková, Pavla (BTO-N)
Cano-Munoz, M. (ES)
Man, Petr (MBU-M) RID, ORCID
Pey, Angel L. (ES)Číslo článku 17200 Zdroj.dok. Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322
Roč. 12, č. 1 (2022)Poč.str. 17 s. Jazyk dok. eng - angličtina Země vyd. DE - Německo Klíč. slova muscle 3-phosphoglycerate kinase ; phosphoglycerate kinase ; phosphoglycerate kinase ; protein stability ; kinetic stability ; molecular-basis ; denaturation ; conformation ; flexibility ; modulation ; ensembles Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology Vědní obor RIV – spolupráce Biotechnologický ústav CEP ED1.1.00/02.0109 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Výzkumná infrastruktura CIISB II - 90127 - Masarykova univerzita Způsob publikování Open access Institucionální podpora MBU-M - RVO:61388971 ; BTO-N - RVO:86652036 UT WOS 000867889200030 EID SCOPUS 85139922175 DOI 10.1038/s41598-022-22088-1 Anotace Phosphoglycerate kinase has been a model for the stability, folding cooperativity and catalysis of a two-domain protein. The human isoform 1 (hPGK1) is associated with cancer development and rare genetic diseases that affect several of its features. To investigate how mutations affect hPGK1 folding landscape and interaction networks, we have introduced mutations at a buried site in the N-terminal domain (F25 mutants) that either created cavities (F25L, F25V, F25A), enhanced conformational entropy (F25G) or introduced structural strain (F25W) and evaluated their effects using biophysical experimental and theoretical methods. All F25 mutants folded well, but showed reduced unfolding cooperativity, kinetic stability and altered activation energetics according to the results from thermal and chemical denaturation analyses. These alterations correlated well with the structural perturbation caused by mutations in the N-terminal domain and the destabilization caused in the interdomain interface as revealed by H/D exchange under native conditions. Importantly, experimental and theoretical analyses showed that these effects are significant even when the perturbation is mild and local. Our approach will be useful to establish the molecular basis of hPGK1 genotype-phenotype correlations due to phosphorylation events and single amino acid substitutions associated with disease. Pracoviště Mikrobiologický ústav Kontakt Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Rok sběru 2023 Elektronická adresa https://www.nature.com/articles/s41598-022-22088-1
Počet záznamů: 1