Case Number 107054 - Novel End-Point Assay for Autophagy

Contact: Ellen Monson
Email: geoffrey.pinski@uc.edu
Phone: 513-558-5274

Description:  Dr. Patrick Dennis at the University of Cincinnati developed a novel end-point assay for autophagy based on the observation that the BHMT (betaine homocysteine methyl transferase) enzyme is degraded via autophagy with the generation of a discrete proteolytic fragment in the lysosomal compartment (1). Dr. Dennis engineered a novel Glutathione-S-Transferase (GST)-BHMT reporter with targeted mutations that result in degradation of the fusion protein via the autophagocytic pathway. Autophagocytic degradation of the reporter leads to the generation of discrete proteolytic fragments in the lysosomal compartment, similar to native BHMT. The advantages of this autophagy assay over existing assays are 1) end-point assay rather than mid-point assay, 2) increased sensitivity and 3) less subjective and more quantifiable.