Abstract
The tendency for tetramerization is the main disadvantage in the green fluorescent protein homologues from Anthozoa species. We report a universal method called hetero-oligomeric tagging, which diminishes troublesome consequences of tetramerizafion of Anthozoa-derived fluorescent proteins (FP) in intracellular protein labelling. This approach is based on the co-expression of the FP-tagged protein of interest together with an excess of free non-fluorescent FP mutant. The resulting FP heterotetramers contain only a single target polypeptide and, therefore, can be considered pseudo-monomeric. Feasibility of the method has been demonstrated with a red FP fused with cytoplasmic β-actin or tubulin-binding protein Tau34. In addition, heterotetramers appeared to be a unique model for biophysical characterization of Anthozoa FPs in pseudo-monomeric state.
Original language | English (US) |
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Pages (from-to) | 109-114 |
Number of pages | 6 |
Journal | Biochemical Journal |
Volume | 371 |
Issue number | 1 |
DOIs | |
State | Published - Apr 1 2003 |
Externally published | Yes |
Keywords
- Coral fluorescent protein
- Green fluorescent protein
- Heterotetramer
- Protein labelling
- Red fluorescent protein
- Tetramerization
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology