Towards determining the structure of human KIFC1, a potential target for drug development in cancer therapy

Normal cell division is initiated upon centrosome duplication and the two centrosomes move towards the cell periphery to form the poles of a bipolar mitotic spindle, a function that is essential for accurate chromosome segregation. Aberrant centrosomes duplication leading to more than two centrosomes is referred to as centrosome amplification. Centrosome amplification causes genetic instability and is a hallmark in cancer cells. Cancer cells survive the multiple mitotic spindle formation by clustering Microtubules Organizing Centres (MTOCs) to form two supernumerary centrosomes and a single pseudo-bipolar spindle during metaphase. KIFC1, a kinesin essential for this process, is particularly interesting as a potential therapeutic target. This is due to its non-essential role in cell division in normal cells but crucial centrosome clustering function in cancer cells with supernumerary centrosomes. The overall goal of this project was to determine the structure of the catalytic domain, also known as motor domain, of KIFC1 to aid future structure-based drug-design. In this project, I have established the expression and purification protocol for the KIFC1 motor domain. The protocol enables to obtain large amounts of protein at highest purity. Although I obtained crystals initially, their reproduction was difficult and therefore high-resolution crystals of the KIFC1 motor domain proved difficult to obtain. As such, the search for crystallisation conditions that allow the formation of high-resolution and reproducible crystals of the KIFC1 motor domain continues. Once this bottleneck is surpassed, it will be possible to use the structure for future structure-based drug-design.