Click Chemistry in Generation of Mammalian Acetylome to Study Shwachman-Diamond Syndrome
MetadataShow full item record
Shwachman Diamond syndrome is an autosomal recessive disease associated with pancreatic insufficiency, skeletal abnormalities, and bone marrow failure. The de Figueiredo lab has recently shown physical and genetic interactions between Sdo1p/SBDS proteins and class I and II histone deacetylases (HDACs) in yeast and mammalian cells, respectively. This study seeks to implement new methods to generate an acetylome to further elucidate the role of acetylation in the function of SBDS, hoping that more information will lead to the development of more effective treatments for those with SDS. Metabolic labeling with click chemistry is a new method that can be used for this purpose. To study if this chemistry is suitable K-562, an erythroleukemia type of mammalian cells, were used as the model organism. The cells were treated with Trichostatin A (TSA), an HDAC inhibitor, to simulate the interaction between SBDS and HDACs. The proteins of these cells were metabolically labeled with sodium 4-pentynoate. These proteins were then isolated and visualized using the integration of click reaction into a horseradish peroxidase (HRP) labeling system. Since no bands were visualized in the 17 kDa range (the size of HDACs), this data does not support the use of click chemistry in creating an acetylome to further elucidate the cellular mechanism of SDS. Though the concentrations recommended in a previous study did not work for this system, generating 4-pentynoate dosage response data may allow for this strategy to work.
Sibley, Robert (2011). Click Chemistry in Generation of Mammalian Acetylome to Study Shwachman-Diamond Syndrome. Texas A&M University. Available electronically from