Coupling tandem affinity purification and quantitative tyrosine iodination to determine subunit stoichiometry of protein complexes
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UMass Chan Affiliations
Program in Gene Function and ExpressionProgram in Molecular Medicine
Graduate School of Biomedical Sciences
Document Type
Journal ArticlePublication Date
2003-08-02Keywords
Electrophoresis, Polyacrylamide Gel; Fungal Proteins; Indicators and Reagents; Iodine Radioisotopes; Kinetics; Polymerase Chain Reaction; Radioisotope Dilution Technique; Tyrosine; YeastsLife Sciences
Medicine and Health Sciences
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Show full item recordAbstract
Rapid protein purification methodologies, such as strategies involving the tandem affinity purification module, have resulted in the identification of a tremendous number of multisubunit protein complexes. Furthermore, in this modern genomic age, mass spectrometry methods are often coupled with affinity purification to identify the genes that encode each protein subunit. However, simple methodologies to determine the stoichiometry of individual subunits within a multisubunit complex have not received much attention. In this article we describe a procedure to rapidly and efficiently determine the stoichiometry of subunits within multisubunit complexes using a combination of tandem affinity purification and quantitative 125I labeling of subunit tyrosines.Source
Methods. 2003 Sep;31(1):104-9.
DOI
10.1016/S1046-2023(03)00094-XPermanent Link to this Item
http://hdl.handle.net/20.500.14038/32565PubMed ID
12893180Related Resources
ae974a485f413a2113503eed53cd6c53
10.1016/S1046-2023(03)00094-X