Label Transfer Chemistry for the Characterization of Protein−Protein Interactions
- 26 September 2007
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 129 (41) , 12348-12349
- https://doi.org/10.1021/ja072904r
Abstract
A new label transfer method is presented that overcomes most of the limitations of current systems. A protein of interest is tagged with a tetracysteine sequence (FlAsH receptor peptide (FRP)) that binds tightly and specifically to a chimeric molecule 3,4-dihydroxyphenylalanine-biotin-4‘,5‘-bis(1,3,2-dithioarsolan-2-yl)fluorescein (DOPA-biotin-FlAsH). Upon brief periodate oxidation, the DOPA moiety is cross-linked to nearby surface-exposed nucleophiles. Boiling the products in excess dithiol dissolves the FlAsH-FRP interaction, resulting in transfer of the biotin tag to the partner proteins, allowing them to be identified by standard methods.Keywords
This publication has 18 references indexed in Scilit:
- Chemistry of Periodate-Mediated Cross-Linking of 3,4-Dihydroxylphenylalanine-Containing Molecules to ProteinsJournal of the American Chemical Society, 2006
- Identification of Gal4 activation domain-binding proteins in the 26S proteasome by periodate-triggered cross-linkingMolecular BioSystems, 2005
- Role of Rpn11 Metalloprotease in Deubiquitination and Degradation by the 26 S ProteasomeScience, 2002
- Recruitment of a 19 S Proteasome Subcomplex to an Activated PromoterScience, 2002
- A proteasomal ATPase subunit recognizes the polyubiquitin degradation signalNature, 2002
- Geobacter metallireducens accesses insoluble Fe(iii) oxide by chemotaxisNature, 2002
- Isolation and Characterization of SUG2Journal of Biological Chemistry, 1996
- Mediation of Epstein-Barr Virus EBNA2 Transactivation by Recombination Signal-Binding Protein J κScience, 1994
- Identification of the Target of a Transcription Activator Protein by Protein-Protein PhotocrosslinkingScience, 1994
- GAL4-VP16 is an unusually potent transcriptional activatorNature, 1988