MOLECULAR CHARACTERISATION OF INOSITOL MONOPHOSPHATASE LIKE (IMP-L) PROTEINS IN CHICKPEA

Abstract

Inositol holds great importance for the physiological well being of organisms. In eukaryotes especially, the intracellular pools of inositol are essential metabolites as well as labile messengers that regulate cellular physiology while travelling within and between cells. Free inositol is synthesised by either the de novo biosynthetic pathway or recycling of inositol phosphates by de-phosphorylation. It is a multifaceted compound that generates myriad of derivatives upon phosphorylation. The key enzyme needed for the myo-inositol biosynthesis functionally, a hydrolase, is known as myo-inositol monophosphatase (IMP). In addition to IMP, two other genes have been hypothesised to have IMP activity that has resulted in their classification as IMP-like (IMPL) proteins. The IMP together with IMPL constitutes the inositol monophosphatase gene family. Inositol monophosphatase (IMP) gene has previously been characterised in chickpea. In this project, we aim to isolate and clone the gene encoding IMPL1 and IMPL2 and its corresponding cDNA from chickpea. Also, we intend to study the bioactivity of the proteins using bacterially over-expressed and purified protein fractions. Further, we also aim to functionally characterise CaIMPL1 and CaIMPL2.