Congenital glycosylation disorders (CDG) are a rare disease group that affect all parts of a patient’s body by disrupting the building of sugar chains (called glycans) in cells. Glycans are used to maintain communication and give structural support between cells of the body. They are made in a specialized part of the cell called the Golgi. We have developed a method for studying outside the cell how the Golgi organizes enzymes to build correct glycans. In the proposed work we will use this method to investigate disease causing mutations in two genes known to cause CDG. A further aim of the work will be to improve our method to follow Golgi organization with much higher precision using a more sophisticated imaging system called Total Internal Reflection Fluorescence Microscopy (TIRFM).
1. Generated a ATPaseV0A2 knock-out reporter cell line, isolated and assayed Golgi from the KO cells.
2. Investigated cytosol from a new Cog1-deficient patient in the cell free assay.
3. Established the collaborations intitiated during proposal writing with the groups of Kronak (ATPase strand, Berlin) and Morava (Cog1 strand, Leuven)