Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation
Tuberculosis (TB) remains one of the major health concerns worldwide. <i>Mycobacterium tuberculosis</i> (Mtb), the causative agent of TB, can flexibly change its metabolic processes during different life stages. Regulation of key metabolic enzyme activities by intracellular conditions, a...
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MDPI AG
2021-02-01
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| Series: | International Journal of Molecular Sciences |
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| author | Jan Snášel Iva Machová Veronika Šolínová Václav Kašička Marcela Krečmerová Iva Pichová |
| author_facet | Jan Snášel Iva Machová Veronika Šolínová Václav Kašička Marcela Krečmerová Iva Pichová |
| author_sort | Jan Snášel |
| collection | DOAJ |
| description | Tuberculosis (TB) remains one of the major health concerns worldwide. <i>Mycobacterium tuberculosis</i> (Mtb), the causative agent of TB, can flexibly change its metabolic processes during different life stages. Regulation of key metabolic enzyme activities by intracellular conditions, allosteric inhibition or feedback control can effectively contribute to Mtb survival under different conditions. Phosphofructokinase (Pfk) is one of the key enzymes regulating glycolysis. Mtb encodes two Pfk isoenzymes, Pfk A/Rv3010c and Pfk B/Rv2029c, which are differently expressed upon transition to the hypoxia-induced non-replicating state of the bacteria. While <i>pfkB</i> gene and protein expression are upregulated under hypoxic conditions, Pfk A levels decrease. Here, we present biochemical characterization of both Pfk isoenzymes, revealing that Pfk A and Pfk B display different kinetic properties. Although the glycolytic activity of Pfk A is higher than that of Pfk B, it is markedly inhibited by an excess of both substrates (fructose-6-phosphate and ATP), reaction products (fructose-1,6-bisphosphate and ADP) and common metabolic allosteric regulators. In contrast, synthesis of fructose-1,6-bisphosphatase catalyzed by Pfk B is not regulated by higher levels of substrates, and metabolites. Importantly, we found that only Pfk B can catalyze the reverse gluconeogenic reaction. Pfk B thus can support glycolysis under conditions inhibiting Pfk A function. |
| first_indexed | 2024-03-09T06:05:18Z |
| format | Article |
| id | doaj.art-586832f9d6a340d09db9e875a967906b |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T06:05:18Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-586832f9d6a340d09db9e875a967906b2023-12-03T12:04:58ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-02-01223148310.3390/ijms22031483Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric RegulationJan Snášel0Iva Machová1Veronika Šolínová2Václav Kašička3Marcela Krečmerová4Iva Pichová5Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, 16610 Prague 6, Czech RepublicInstitute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, 16610 Prague 6, Czech RepublicInstitute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, 16610 Prague 6, Czech RepublicInstitute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, 16610 Prague 6, Czech RepublicInstitute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, 16610 Prague 6, Czech RepublicInstitute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo nám. 2, 16610 Prague 6, Czech RepublicTuberculosis (TB) remains one of the major health concerns worldwide. <i>Mycobacterium tuberculosis</i> (Mtb), the causative agent of TB, can flexibly change its metabolic processes during different life stages. Regulation of key metabolic enzyme activities by intracellular conditions, allosteric inhibition or feedback control can effectively contribute to Mtb survival under different conditions. Phosphofructokinase (Pfk) is one of the key enzymes regulating glycolysis. Mtb encodes two Pfk isoenzymes, Pfk A/Rv3010c and Pfk B/Rv2029c, which are differently expressed upon transition to the hypoxia-induced non-replicating state of the bacteria. While <i>pfkB</i> gene and protein expression are upregulated under hypoxic conditions, Pfk A levels decrease. Here, we present biochemical characterization of both Pfk isoenzymes, revealing that Pfk A and Pfk B display different kinetic properties. Although the glycolytic activity of Pfk A is higher than that of Pfk B, it is markedly inhibited by an excess of both substrates (fructose-6-phosphate and ATP), reaction products (fructose-1,6-bisphosphate and ADP) and common metabolic allosteric regulators. In contrast, synthesis of fructose-1,6-bisphosphatase catalyzed by Pfk B is not regulated by higher levels of substrates, and metabolites. Importantly, we found that only Pfk B can catalyze the reverse gluconeogenic reaction. Pfk B thus can support glycolysis under conditions inhibiting Pfk A function.https://www.mdpi.com/1422-0067/22/3/1483<i>Mycobacterium tuberculosis</i>glycolysisphosphofructokinase A and Ballosteric regulationenzyme kinetics |
| spellingShingle | Jan Snášel Iva Machová Veronika Šolínová Václav Kašička Marcela Krečmerová Iva Pichová Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation International Journal of Molecular Sciences <i>Mycobacterium tuberculosis</i> glycolysis phosphofructokinase A and B allosteric regulation enzyme kinetics |
| title | Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation |
| title_full | Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation |
| title_fullStr | Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation |
| title_full_unstemmed | Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation |
| title_short | Phosphofructokinases A and B from <i>Mycobacterium tuberculosis</i> Display Different Catalytic Properties and Allosteric Regulation |
| title_sort | phosphofructokinases a and b from i mycobacterium tuberculosis i display different catalytic properties and allosteric regulation |
| topic | <i>Mycobacterium tuberculosis</i> glycolysis phosphofructokinase A and B allosteric regulation enzyme kinetics |
| url | https://www.mdpi.com/1422-0067/22/3/1483 |
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