In my everyday life, I try to balance and time manage all the various activities I am involved with. Apart from life in the lab, I have a passion for football. In fact I train almost everyday with a local football team. I also have a passion for video games, mainly Assassin's Creed games. I love to go out out and enjoy the company of my girlfriend, family and friends. I aspire to continue my studies at a PhD level and continue to leave an impact on society from behind the scenes.
Functional Effects of Glucokinase Variants Identified in Diabetic Patients in Malta
My current research involves the pancreatic glucokinase protein that functions as an enzyme to convert glucose into glucose-6-phosphate in the glycolysis pathway. Mutations, identified in a Maltese cohort that suffer from diabetes, were generated within the gene that codes for this protein through site-directed mutagenesis. In silico cloning was carried out followed by in vitro expression and purification of the wildtype and mutant glucokinase proteins. E. coli competent cells were transformed with clones, consisting of a vector and the glucokinase gene insert and optimization of conditions was carried out. Proteins were purified using Immobilized Metal Ion Affinity Chromatography. Proteins were characterized to check whether the function, stability and structure changed due to the mutations. Native-PAGE analysis, circular dichroism, activity assays, isothermal titration calorimetry, spectrophotometric enzymatic assays are being carried out on the protein samples.
The Next step…
Future research will include crystallization of wildtype and mutant protein samples to analyze the crystal structure and determine the specific changes caused by the mutations. Not all mutations discovered in the Maltese cohort were used and characterized in this study thus further work can be carried out to analyze the remaining mutations.