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Existing variants of green fluorescent protein GFP often misfold when expressed as fusions with other proteins. We have generated a robustly folded version of GFP, called 'superfolder' GFP, that folds well even when fused to poorly folded polypeptides.
The fluorescence of Escherichia coli cells expressing each of eighteen proteins from Pyrobaculum aerophilum as fusions with superfolder GFP was proportional to total protein expression. In contrast, fluorescence of folding reporter GFP fusion proteins was strongly correlated with the productive folding yield of the passenger protein. X-ray crystallographic structural analyses helped explain the enhanced folding of superfolder GFP relative to folding reporter GFP.
Engineering and characterization of a superfolder green fluorescent protein. Nat Biotechnol 24 Cited: times. PMID: Abstract Existing variants of green fluorescent protein GFP often misfold when expressed as fusions with other proteins.
Front Mol Biosci 8 Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting. Sci Rep 6 Conformational changes of the HIV-1 envelope protein during membrane fusion are inhibited by the replacement of its membrane-spanning domain. J Biol Chem Approaching infinite affinity through engineering of peptide-protein interaction.