Boston Biomedical Research Institute

3 ARTICLES PUBLISHED IN JoVE

Biology

Growth Assays to Assess Polyglutamine Toxicity in Yeast

This manuscript describes three complementary protocols for assessing the toxicity of polyglutamine (polyQ)-expansion proteins in the yeast Saccharomyces cerevisiae. These protocols can easily be modified to monitor the toxicity of other misfolded proteins in yeast.

Biology

Protease- and Acid-catalyzed Labeling Workflows Employing ¹⁸O-enriched Water

Diana Klingler¹, Markus Hardt¹

¹Boston Biomedical Research Institute

Stable isotope labeling workflows employing ¹⁸O-enriched water (LeO-workflows) are versatile tools for quantitative and qualitative proteomics studies. In protease-assisted (PALeO) workflows, ¹⁸O-atoms are introduced by proteolytic cleavage and carboxyl oxygen exchange reactions mediated by proteases. In the acid-catalyzed (ALeO) workflow, ¹⁸O-atoms are introduced by carboxyl oxygen exchange at low pH.

Biology

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Yuwei Jiang¹, Sonja Di Gregorio², Maram B. Albakri¹, Martin L. Duennwald^1,2, Patrick Lajoie¹

¹Department of Anatomy and Cell Biology, University of Western Ontario, ²Department of Pathology and Laboratory Medicine, University of Western Ontario

This article describes protocols to assess the effect of fluorescent proteins on the aggregation and toxicity of misfolded polyglutamine expansion for the rapid evaluation of a newly uncharacterized fluorescent protein in the context of fluorescent reporters.