title: A rewired green fluorescent protein: folding and function in a nonsequential, noncircular GFP permutant.
abstract: The sequential order of secondary structural elements in proteins affects the folding and activity to an unknown extent. To test the dependence on sequential connectivity, we reconnected secondary structural elements by their solvent-exposed ends, permuting their sequential order, called "rewiring". This new protein design strategy changes the topology of the backbone without changing the core side chain packing arrangement. While circular and noncircular permutations have been observed in protein structures that are not related by sequence homology, to date no one has attempted to rationally design and construct a protein with a sequence that is noncircularly permuted while conserving three-dimensional structure. Herein, we show that green fluorescent protein can be rewired, still functionally fold, and exhibit wild-type fluorescence excitation and emission spectra.
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Philippa J
authors.initials: PJ
authors.lastname: Reeder
authors.forename: Yao-Ming
authors.initials: YM
authors.lastname: Huang
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
authors.forename: Christopher
authors.initials: C
authors.lastname: Bystroff
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D003063:Cnidaria / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics*; D009154:Mutation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins