Počet záznamů: 1
The Role of Cysteine Residues in Catalysis of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis
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SYSNO ASEP 0474974 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název The Role of Cysteine Residues in Catalysis of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis Tvůrce(i) Machová, Iva (UOCHB-X) RID, ORCID
Hubálek, Martin (UOCHB-X) RID, ORCID
Lepšík, Martin (UOCHB-X) RID, ORCID
Bednárová, Lucie (UOCHB-X) RID, ORCID
Pazderková, Markéta (UOCHB-X) RID, ORCID
Kopecký, V. Jr. (CZ)
Snášel, Jan (UOCHB-X) RID
Dostál, Jiří (UOCHB-X) RID, ORCID
Pichová, Iva (UOCHB-X) RID, ORCIDČíslo článku e0170373 Zdroj.dok. PLoS ONE. - : Public Library of Science - ISSN 1932-6203
Roč. 12, č. 1 (2017)Poč.str. 12 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova structural insights ; metabolism ; parameters Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology CEP LO1302 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy GBP208/12/G016 GA ČR - Grantová agentura ČR Institucionální podpora UOCHB-X - RVO:61388963 UT WOS 000396124700012 EID SCOPUS 85011298587 DOI 10.1371/journal.pone.0170373 Anotace Mycobacterium tuberculosis (MTb), the causative agent of tuberculosis, can persist in macrophages for decades, maintaining its basic metabolic activities. Phosphoenolpyruvate carboxykinase (Pck, EC 4.1.1.32) is a key player in central carbon metabolism regulation. In replicating MTb, Pck is associated with gluconeogenesis, but in non-replicating MTb, it also catalyzes the reverse anaplerotic reaction. Here, we explored the role of selected cysteine residues in function of MTb Pck under different redox conditions. Using mass spectrometry analysis we confirmed formation of S-S bridge between cysteines C391 and C397 localized in the C-terminal subdomain. Molecular dynamics simulations of C391-C397 bridged model indicated local conformation changes needed for formation of the disulfide. Further, we used circular dichroism and Raman spectroscopy to analyze the influence of C391 and C397 mutations on Pck secondary and tertiary structures, and on enzyme activity and specificity. We demonstrate the regulatory role of C391 and C397 that form the S-S bridge and in the reduced form stabilize Pck tertiary structure and conformation for gluconeogenic and anaplerotic reactions. Pracoviště Ústav organické chemie a biochemie Kontakt asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Rok sběru 2018 Elektronická adresa http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0170373
Počet záznamů: 1