Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Cycloheximide / pharmacology
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Cysteine Endopeptidases / metabolism
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Fungal Proteins / metabolism
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Half-Life
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HeLa Cells
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Humans
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Immunoblotting
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Isotope Labeling
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Methods
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Multienzyme Complexes / metabolism
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Promoter Regions, Genetic / drug effects
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Promoter Regions, Genetic / genetics
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Proteasome Endopeptidase Complex
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Protein Processing, Post-Translational*
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Protein Synthesis Inhibitors / pharmacology
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Proteins / metabolism*
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Yeasts / metabolism
Substances
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Fungal Proteins
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Multienzyme Complexes
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Protein Synthesis Inhibitors
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Proteins
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Cycloheximide
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Cysteine Endopeptidases
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Proteasome Endopeptidase Complex