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Quantifying the activation of the small molecular weight G‐protein ras
Author(s) -
Chapman Katharine,
Hall David J.
Publication year - 2005
Publication title -
biochemistry and molecular biology education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 39
eISSN - 1539-3429
pISSN - 1470-8175
DOI - 10.1002/bmb.2005.494033010420
Subject(s) - gtp' , guanine nucleotide exchange factor , lysis , chemistry , biochemistry , guanine , microbiology and biotechnology , kinase , protein kinase a , g protein , receptor , ligand (biochemistry) , gtp binding protein regulators , gtpase , nucleotide , biology , enzyme , gene
Low molecular weight guanine‐nucleotide binding proteins are molecular switches involved in a wide range of cellular functions in eukaryotes. Ras is activated when a membrane receptor is bound by a ligand and the GDP bound to the Ras protein is exchanged for GTP. Once GTP is bound, the Ras three‐dimensional conformation changes and binds the kinase Raf. This protein‐protein interaction can be used to “pull‐down” activated Ras from a cell lysate. The amount of activated Ras is quantitated through immunoblotting and densitometry. This procedure is excellent as a multiweek investigational laboratory.