Determining protein half-lives.
Controlling the stability of cellular proteins is a fundamental way by which cells regulate growth, differentiation, survival, and development. Measuring the turnover rate of a protein is often the first step in assessing whether or not the function of a protein is regulated by proteolysis under specific physiological conditions. Over the years, procedures to determine the half-life of proteins in cultured eukaryotic cells have been well-established. This chapter describes in detail the two most frequently used methods, pulse-chase analysis and cycloheximide blocking, to determine a protein's half-life in yeast and cultured mammalian cells.
Zhou
Pengbo
P
Department of Pathology and Laboratory Medicine, Weill Medical College of Cornell University, New York, NY, USA.
D003513:Cycloheximide / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D006207:Half-Life; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D007553:Isotope Labeling; D008722:Methods; D009097:Multienzyme Complexes / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D046988:Proteasome Endopeptidase Complex; D011499:Protein Processing, Post-Translational; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism*; D015003:Yeasts / Q000378:metabolism
D005656:Fungal Proteins; D009097:Multienzyme Complexes; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D003513:Cycloheximide; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex