Novel Substrates for Kinases Involved in the Biosynthesis of Inositol Pyrophosphates and Their Enhancement of ATPase Activity of a Kinase

• Published in: Molecules (Basel, Switzerland) Vol. 26; no. 12; p. 3601
• Main Authors: Mohanrao, Raja, Manorama, Ruth, Ganguli, Shubhra, Madhusudhanan, Mithun C., Bhandari, Rashna, Sureshan, Kana M.
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 11-06-2021; MDPI
• Subjects: Adenosine triphosphatase; ATP; ATPase; Biosynthesis; Enzymatic activity; Enzyme activity; Enzymes; Inositol phosphate; Inositol phosphates; inositol pyrophosphate; IP6K1; kinase; Kinases; Phosphates; Phosphorylation; Polyphosphates; Substrate specificity; Substrates; VIP1 KD; Yeasts
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules26123601

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 scyllo-inositol polyphosphates—scyllo-IP5, scyllo-IP6, scyllo-IP7 and Bz-scyllo-IP5—from myo-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 scyllo-inositol polyphosphates with these kinases and ATP as the phosphate donor. We tracked enzyme activity by measuring the amount of radiolabeled scyllo-inositol pyrophosphate product formed and the amount of ATP consumed. All scyllo-inositol polyphosphates are substrates for both the kinases but they are weaker than the corresponding myo-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-scyllo-IP5 was the most potent ligand to induce IP6K1 ATPase activity despite being a weak substrate. This compound could have potential as a competitive inhibitor.