Membrane proteins (MPs) play a major role in a wide range of biological processes and represent one third of the total proteins encoded by the human genome. Among these, ion channels and transporters play a basic role in pain-causing stimuli (TRP channels), epithelial chloride transport in many organs (CFTR), c-type inactivation (voltage gated potassium channels), neurotransmitter release and regulating contractile tone (BK channels), mitochondrial-membrane transport (VDAC), in the absorption of dietary peptides (PEPT1 transporter), hepatic uptake of drugs and small molecules (OCT1), drug absorption and disposition (PGP1 or MDR1), glucose transport (SGLT1), regulating excitatory neurotransmitter glutamate at synapses (EAAT). Any functional defect in the MPs could affect the cellular activities which could often lead to a wide range of diseases like cystic fibrosis, cancer, pain, autoimmune diseases, stroke, apoptosis, intestinal inflammation, glucose-galactose malabsorption, hepatic steatosis, amyotrophic lateral sclerosis (ALS), unexpected toxicities or drug-drug interactions etc. Cell-free protein synthesis (CFPS) is a versatile, flexible, faster and alternative approach for the synthesis of difficult-to-express proteins including MPs and toxic proteins. CFPS has a high degree of controllability and provides a completely open system allowing direct manipulation of the reaction conditions to optimize protein folding, disulfide bond formation, and incorporation of noncanonical amino acids. One can add the detergents, liposomes or make use of the native endogenous microsomes for the solubilization and functional folding of the MPs. Thus, efficient use of CFPS helps in the economic production of functional MPs as targets for drug development.
The goal of this project is to synthesize ion channels and transporters using cell-free systems based on eukaryotic and prokaryotic extracts and develop functional and pharmacological assays. The main focus will be on the investigation of clinically significant proteins (mentioned above).
- Good academic records with a Master’s degree or Diploma in Pharmacology, Pharmacy, Biophysics, or Biochemistry.
- Candidates with experience or interest in liposomes, electrophysiology, and functional characterization of membrane proteins will be favored.
- Applicants should have excellent English language skills.