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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 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Loss of stability and unfolding cooperativity in hPGK1 upon gradual structural perturbation of its N-terminal domain hydrophobic core Author(s) Luis Pacheco-Garcia, J. (ES)
Loginov, Dmitry Sergej (MBU-M) RID, ORCID
Naganathan, A. N. (IN)
Vaňková, Pavla (BTO-N) RID, ORCID
Cano-Munoz, M. (ES)
Man, Petr (MBU-M) RID, ORCID
Pey, Angel L. (ES)Article number 17200 Source Title Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322
Roč. 12, č. 1 (2022)Number of pages 17 s. Language eng - English Country DE - Germany Keywords muscle 3-phosphoglycerate kinase ; phosphoglycerate kinase ; phosphoglycerate kinase ; protein stability ; kinetic stability ; molecular-basis ; denaturation ; conformation ; flexibility ; modulation ; ensembles Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology Subject RIV - cooperation Institute of Biotechnology R&D Projects ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Research Infrastructure CIISB II - 90127 - Masarykova univerzita Method of publishing Open access Institutional support MBU-M - RVO:61388971 ; BTO-N - RVO:86652036 UT WOS 000867889200030 EID SCOPUS 85139922175 DOI https://doi.org/10.1038/s41598-022-22088-1 Annotation 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. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2023 Electronic address https://www.nature.com/articles/s41598-022-22088-1
Number of the records: 1