Influenza is an infectious disease responsible for severe illness and death, and capable of worldwide pandemics.
Current treatment is limited to the use of M2 ion channel inhibitors (amantadine and remantadine) and inhibitors of influenza neuraminidase (oseltamivir and zanamivir). Globally, an increase in drug resistance to standard anti-influenza mediciations has been observed. Therefore, an urgent need to develop new drugs to combat the symptoms and spread of this disease has become evident.
The Krause lab is involved in the development of novel neuraminidase inhibitors. Neuraminidase is an essential protein involved in the release of progeny virus particles from cells and the movement of the virus through the respiratory tract. When inhibited, viral spread between hosts is limited and symptoms of the disease decrease rapidly. In a collaboration with scientists working in J. Andrew McCammon's group at UCSD, a set of 27 new inhibitors have been indentified in silico using molecular dynamic simulations, which have predicted binding values equal to or greater than current neuraminidase inhibitors. These inhibitors target a unique cavity in the protein and a major goal is to test these inhibitors in vitro and explain the molecular basis of this targeting.
The neuraminidase project involves the cloning and expression of the neuraminidase protein for biophysical characterisation of these potential influenza drugs. This involves experimental procedures including DNA cloning procedures, optimisation of neuraminidase expression in insect and mammalian cell lines, optimisation of neuraminidase purification, a fluorometric assay for neuraminidase inhibition, and biophysical analysis including isothermal calorimetry, dynamic light scattering and protein crystallisation.