Publication Abstract from PubMed
Bimolecular fluorescence complementation (BiFC) assay has been used widely to visualize protein-protein interactions in cells. However, there is a problem that fluorescent protein fragments have an ability to associate with each other independent of an interaction between proteins fused to the fragments. To facilitate the BiFC assay, we have attempted to determine the structure and characteristics of reassembled fluorescent protein, Venus. The anion-exchange chromatography showed an oligomer and a monomer of reassembled Venus. Our results suggested that the oligomer was formed by beta-strands swapping without any serious steric clashes and was converted to the monomer. Crystal structure of reassembled Venus had an 11-stranded beta-barrel fold, typical of GFP-derived fluorescent proteins. Based on the structural features, we have mutated to beta-strand 7 and measured T(m) values. The results have revealed that the mutation influences the thermal stability of reassembled fluorescent complex.
Structure and characteristics of reassembled fluorescent protein, a new insight into the reassembly mechanisms.,Isogai M, Kawamoto Y, Inahata K, Fukada H, Sugimoto K, Tada T Bioorg Med Chem Lett. 2011 May 15;21(10):3021-4. Epub 2011 Mar 16. PMID:21463942[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
