The greatest bottleneck in structural studies of Sacsin is the preparation and screening of fragments of thousands of Sacsin constructs with different boundaries, different mutations, and sequences from different species. To overcome this, we have developed a high-throughput strategy that relies on chemical DNA synthesis to prepare medium-sized libraries (mixtures) of Sacsin protein fragments. These are analyzed by gel filtration to isolate well-behaved proteins with solubility and stability characteristics that are favourable for structural studies. The proteins with these properties are then identified by mass spectrometry and studied by structural biology. The goal of the high–throughput strategy is to identify well-behaved Sacsin protein fragments for structural studies.
Duration: one year
Grant: $98,280
Contact:
Dr Kalle Gehring
Dr. Kalle Gehring, Professor Department of Biochemistry MCGill University
Francesco Bellini Life Sciences Building, 3649 promenade Sir-William-Osler, Office: Room 469; Lab: Room 473, Montreal, Quebec H3G 0B1
Tel:(514)-398-7287; Lab: (514)-398-2873/1496; Fax:(514)-398-2983; Email: kalle.gehring@mcgill.ca