A novel multiple affinity purification tag and its use in identification of proteins associated with a cyclin-CDK complex
- 15 February 2001
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 29 (4) , 24e-24
- https://doi.org/10.1093/nar/29.4.e24
Abstract
A novel multiple affinity purification (MAFT) or tandem affinity purification (TAP) tag has been constructed. It consists of the calmodulin binding peptide, six histidine residues, and three copies of the hemagglutinin epitope. This 'CHH' MAFT tag allows two or three consecutive purification steps, giving high purity. Active Clb2-Cdc28 kinase complex was purified from yeast cells after inserting the CHH tag into Clb2. Associated proteins were identified using mass spectrometry. These included the known associated proteins Cdc28, Sic1 and Cks1. Several other proteins were found including the 70 kDa chaperone, Ssa1.Keywords
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