Hetero-oligomeric tagging diminishes non-specific aggregation of target proteins fused with Anthozoa fluorescent proteins

Maria E. Bulina, Vladistav V. Verkhusha, Dmitry B. Staroverov, Dmitry M. Chudakov, Konstantin A. Lukyanov

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

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 languageEnglish (US)
Pages (from-to)109-114
Number of pages6
JournalBiochemical Journal
Volume371
Issue number1
DOIs
StatePublished - Apr 1 2003

Keywords

  • Coral fluorescent protein
  • Green fluorescent protein
  • Heterotetramer
  • Protein labelling
  • Red fluorescent protein
  • Tetramerization

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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