ArticleNucleic acids research2006
Protein-protein interaction analysis by C-terminally specific fluorescence labeling and fluorescence cross-correlation spectroscopy.
Article in Nucleic acids research, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Article
- The Thermodynamic Fingerprints of Ultra-Tight Nanobody-Antigen Binding Probed via Two-Color Single-Molecule Coincidence Detection.International journal of molecular sciences · 2023Article
- A Quantitative Study of Internal and External Interactions of Homodimeric Glucocorticoid Receptor Using Fluorescence Cross-Correlation Spectroscopy in a Live Cell.Scientific reports · 2017Article
- Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance Plasmon Analysis.PloS one · 2015Article
- Dynamics of the ternary complex formed by c-Myc interactor JPO2, transcriptional co-activator LEDGF/p75, and chromatin.The Journal of biological chemistry · 2014Article
- Pulsed interleaved excitation fluctuation imaging.Biophysical journal · 2013Article
- Quantitative analysis of prenylated RhoA interaction with its chaperone, RhoGDI.The Journal of biological chemistry · 2012Article
- Direct observation of dimerization between different CREB1 isoforms in a living cell.PloS one · 2011Article
- Species-independent translational leaders facilitate cell-free expression.Nature biotechnology · 2009Article
- A tetracycline repressor-based mammalian two-hybrid system to detect protein-protein interactions in vivo.Analytical biochemistry · 2009Article
- Fluorescence cross-correlation spectroscopy using single wavelength laser.Frontiers of chemistry in China : selected publications from Chinese universities · 2009Article
- A bacterial two-hybrid system based on the twin-arginine transporter pathway of E. coli.Protein science : a publication of the Protein Society · 2007Article
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Here, we describe novel puromycin derivatives conjugated with iminobiotin and a fluorescent dye that can be linked covalently to the C-terminus of full-length proteins during cell-free translation. The iminobiotin-labeled proteins can be highly purified by affinity purification with streptavidin beads. We confirmed that the purified fluorescence-labeled proteins are useful for quantitative protein-protein interaction analysis based on fluorescence cross-correlation spectroscopy (FCCS). The apparent dissociation constants of model protein pairs such as proto-oncogenes c-Fos/c-Jun and archetypes of the family of Ca2+-modulated calmodulin/related binding proteins were in accordance with the reported values. Further, detailed analysis of the interactions of the components of polycomb group complex, Bmi1, M33, Ring1A and RYBP, was successfully conducted by means of interaction assay for all combinatorial pairs. The results indicate that FCCS analysis with puromycin-based labeling and purification of proteins is effective and convenient for in vitro protein-protein interaction assay, and the method should contribute to a better understanding of protein functions by using the resource of available nucleotide sequences.
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