Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase
IP6K and PPIP5K are two kinases involved in the synthesis of inositol pyrophosphates. Synthetic analogs or mimics are necessary to understand the substrate specificity of these enzymes and to find molecules that can alter inositol pyrophosphate synthesis. In this context, we synthesized four <i&g...
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MDPI AG
2021-06-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/12/3601 |
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| author | Raja Mohanrao Ruth Manorama Shubhra Ganguli Mithun C. Madhusudhanan Rashna Bhandari Kana M. Sureshan |
| author_facet | Raja Mohanrao Ruth Manorama Shubhra Ganguli Mithun C. Madhusudhanan Rashna Bhandari Kana M. Sureshan |
| author_sort | Raja Mohanrao |
| collection | DOAJ |
| description | IP6K and PPIP5K are two kinases involved in the synthesis of inositol pyrophosphates. Synthetic analogs or mimics are necessary to understand the substrate specificity of these enzymes and to find molecules that can alter inositol pyrophosphate synthesis. In this context, we synthesized four <i>scyllo</i>-inositol polyphosphates—<i>scyllo</i>-IP<sub>5</sub>, <i>scyllo</i>-IP<sub>6</sub>, <i>scyllo</i>-IP<sub>7</sub> and Bz-<i>scyllo</i>-IP<sub>5</sub>—from <i>myo</i>-inositol and studied their activity as substrates for mouse IP6K1 and the catalytic domain of VIP1, the budding yeast variant of PPIP5K. We incubated these <i>scyllo</i>-inositol polyphosphates with these kinases and ATP as the phosphate donor. We tracked enzyme activity by measuring the amount of radiolabeled <i>scyllo</i>-inositol pyrophosphate product formed and the amount of ATP consumed. All <i>scyllo</i>-inositol polyphosphates are substrates for both the kinases but they are weaker than the corresponding <i>myo</i>-inositol phosphate. Our study reveals the importance of axial-hydroxyl/phosphate for IP6K1 substrate recognition. We found that all these derivatives enhance the ATPase activity of VIP1. We found very weak ligand-induced ATPase activity for IP6K1. Benzoyl-<i>scyllo</i>-IP<sub>5</sub> was the most potent ligand to induce IP6K1 ATPase activity despite being a weak substrate. This compound could have potential as a competitive inhibitor. |
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| language | English |
| last_indexed | 2024-03-10T10:28:15Z |
| publishDate | 2021-06-01 |
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| series | Molecules |
| spelling | doaj.art-6294e5f1ad47407e8e9499d56afabd9d2023-11-21T23:50:13ZengMDPI AGMolecules1420-30492021-06-012612360110.3390/molecules26123601Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a KinaseRaja Mohanrao0Ruth Manorama1Shubhra Ganguli2Mithun C. Madhusudhanan3Rashna Bhandari4Kana M. Sureshan5School of Chemistry, Indian Institute of Science Education and Research Thiruvananthapuram, Kerala 695551, IndiaLaboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics, Hyderabad 500039, IndiaLaboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics, Hyderabad 500039, IndiaSchool of Chemistry, Indian Institute of Science Education and Research Thiruvananthapuram, Kerala 695551, IndiaLaboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics, Hyderabad 500039, IndiaSchool of Chemistry, Indian Institute of Science Education and Research Thiruvananthapuram, Kerala 695551, IndiaIP6K and PPIP5K are two kinases involved in the synthesis of inositol pyrophosphates. Synthetic analogs or mimics are necessary to understand the substrate specificity of these enzymes and to find molecules that can alter inositol pyrophosphate synthesis. In this context, we synthesized four <i>scyllo</i>-inositol polyphosphates—<i>scyllo</i>-IP<sub>5</sub>, <i>scyllo</i>-IP<sub>6</sub>, <i>scyllo</i>-IP<sub>7</sub> and Bz-<i>scyllo</i>-IP<sub>5</sub>—from <i>myo</i>-inositol and studied their activity as substrates for mouse IP6K1 and the catalytic domain of VIP1, the budding yeast variant of PPIP5K. We incubated these <i>scyllo</i>-inositol polyphosphates with these kinases and ATP as the phosphate donor. We tracked enzyme activity by measuring the amount of radiolabeled <i>scyllo</i>-inositol pyrophosphate product formed and the amount of ATP consumed. All <i>scyllo</i>-inositol polyphosphates are substrates for both the kinases but they are weaker than the corresponding <i>myo</i>-inositol phosphate. Our study reveals the importance of axial-hydroxyl/phosphate for IP6K1 substrate recognition. We found that all these derivatives enhance the ATPase activity of VIP1. We found very weak ligand-induced ATPase activity for IP6K1. Benzoyl-<i>scyllo</i>-IP<sub>5</sub> was the most potent ligand to induce IP6K1 ATPase activity despite being a weak substrate. This compound could have potential as a competitive inhibitor.https://www.mdpi.com/1420-3049/26/12/3601kinaseinositol pyrophosphateIP6K1VIP1 KDATPase |
| spellingShingle | Raja Mohanrao Ruth Manorama Shubhra Ganguli Mithun C. Madhusudhanan Rashna Bhandari Kana M. Sureshan Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase Molecules kinase inositol pyrophosphate IP6K1 VIP1 KD ATPase |
| title | Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase |
| title_full | Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase |
| title_fullStr | Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase |
| title_full_unstemmed | Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase |
| title_short | Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase |
| title_sort | novel substrates for kinases involved in the biosynthesis of inositol pyrophosphates and their enhancement of atpase activity of a kinase |
| topic | kinase inositol pyrophosphate IP6K1 VIP1 KD ATPase |
| url | https://www.mdpi.com/1420-3049/26/12/3601 |
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