Amyotrophic lateral sclerosis (ALS) is a terrible disease that affects the way a person speaks, moves and even breaths, and is responsible for as many as 5 deaths for every 100,000 people over the age of 20. ALS is a progressive disease, affecting the neurons in the both the upper and lower brain way[2], as the nerve cells die, the disease spreads and infiltrates new cells. As to what causes the cells to die, it is believed to be led in part by a mismanagement of autophagy led by a trimeric protein complex composed of C9orf72, SMCR8 and WDR41. C9orf72 represents one of over 20 genes associated with familial ALS and accounts for as many as 40% of familial ALS . C9orf72 and SMCR8 are both DENN domain containing proteins and involved in autophagy regulation. Whereas WDR41 is a WD-repeat protein and is required for the recruitment of C9orf72 and SMCR8 to lysosomes[3].
Our goal is to structurally characterize the C9orf72-SMCR8-WDR41 trimeric complex using Cryo-EM in collaboration with Dr. Laura Diaz Saez from Kilian Huber lab at SGC Oxford. At the same time, we will aim to determine the crystal structure of WDR41 protein independently using X-ray crystallography at SGC Toronto to support drug-discovery efforts targeting WDR41 protein. For this reason, we first tested multiple combinations of the C9orf72-SMCR8-WDR41 trimeric complex Additionally, we have also designed several WDR41 constructs and placed them into a variety of vectors for both E.coli and SF9 insect cells expression systems in order to identify a well expressing and soluble WDR41 protein construct, suitable for crystallization studies.
Next week we will have preliminary results of construct designs, and test expression
[1] https://www.hopkinsmedicine.org/neurology_neurosurgery/centers_clinics/als/conditions/als_amyotrophic_lateral_sclerosis.html
[2] https://www.mayoclinic.org/diseases-conditions/amyotrophic-lateral-sclerosis/symptoms-causes/syc-20354022