Počet záznamů: 1
Structural and Functional Studies of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis
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SYSNO ASEP 0444002 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Structural and Functional Studies of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis Tvůrce(i) Machová, Iva (UOCHB-X) RID, ORCID
Snášel, Jan (UOCHB-X) RID
Dostál, Jiří (UOCHB-X) RID, ORCID
Brynda, Jiří (UOCHB-X) RID, ORCID
Fanfrlík, Jindřich (UOCHB-X) RID, ORCID
Singh, M. (DE)
Tarábek, Ján (UOCHB-X) RID, ORCID
Vaněk, O. (CZ)
Bednárová, Lucie (UOCHB-X) RID, ORCID
Pichová, Iva (UOCHB-X) RID, ORCIDCelkový počet autorů 10 Zdroj.dok. PLoS ONE. - : Public Library of Science - ISSN 1932-6203
Roč. 10, č. 3 (2015), e0120682/1-e0120682/21Poč.str. 21 s. Forma vydání Online - E Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova crystal structure ; noncovalent complexes ; Mycobacterium tuberculosis ; mechanism Vědní obor RIV CE - Biochemie CEP LO1302 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Institucionální podpora UOCHB-X - RVO:61388963 UT WOS 000351987300164 EID SCOPUS 84925740525 DOI 10.1371/journal.pone.0120682 Anotace Tuberculosis, the second leading infectious disease killer after HIV, remains a top public health priority. The causative agent of tuberculosis, Mycobacterium tuberculosis (Mtb), which can cause both acute and clinically latent infections, reprograms metabolism in response to the host niche. Phosphoenolpyruvate carboxykinase (Pck) is the enzyme at the center of the phosphoenolpyruvate-pyruvate-oxaloacetate node, which is involved in regulating the carbon flow distribution to catabolism, anabolism, or respiration in different states of Mtb infection. Under standard growth conditions, Mtb Pck is associated with gluconeogenesis and catalyzes the metal-dependent formation of phosphoenolpyruvate. In non-replicating Mtb, Pck can catalyze anaplerotic biosynthesis of oxaloacetate. Here, we present insights into the regulation of Mtb Pck activity by divalent cations. Through analysis of the Xray structure of Pck-GDP and Pck-GDP-Mn2+ complexes, mutational analysis of the GDP binding site, and quantum mechanical (QM)-based analysis, we explored the structural determinants of efficient Mtb Pck catalysis. We demonstrate that Mtb Pck requires presence of Mn2+ and Mg2+ cations for efficient catalysis of gluconeogenic and anaplerotic reactions. The anaplerotic reaction, which preferably functions in reducing conditions that are characteristic for slowed or stopped Mtb replication, is also effectively activated by Fe2+ in the presence of Mn2+ or Mg2+ cations. In contrast, simultaneous presence of Fe2+ and Mn2+ or Mg2+ inhibits the gluconeogenic reaction. These results suggest that inorganic ions can contribute to regulation of central carbon metabolism by influencing the activity of Pck. Furthermore, the X-ray structure determination, biochemical characterization, and QM analysis of Pck mutants confirmed the important role of the Phe triad for proper binding of the GDP-Mn2+ complex in the nucleotide binding site and efficient catalysis of the anaplerotic reaction. Pracoviště Ústav organické chemie a biochemie Kontakt asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Rok sběru 2016 Elektronická adresa http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0120682
Počet záznamů: 1