A Suite of Engineered GFP Molecules for Oligomeric Scaffolding

Structure. 2015 Sep 1;23(9):1754-1768. doi: 10.1016/j.str.2015.07.008. Epub 2015 Aug 13.


Applications ranging from synthetic biology to protein crystallization could be advanced by facile systems for connecting multiple proteins together in predefined spatial relationships. One approach to this goal is to engineer many distinct assembly forms of a single carrier protein or scaffold, to which other proteins of interest can then be readily attached. In this work we chose GFP as a scaffold and engineered many alternative oligomeric forms, driven by either specific disulfide bond formation or metal ion addition. We generated a wide range of spatial arrangements of GFP subunits from 11 different oligomeric variants, and determined their X-ray structures in a total of 33 distinct crystal forms. Some of the oligomeric GFP variants show geometric polymorphism depending on conditions, while others show considerable geometric rigidity. Potential future applications of this system are discussed.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Crystallography, X-Ray
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / genetics*
  • Models, Molecular
  • Mutagenesis, Site-Directed / methods*
  • Protein Folding
  • Protein Multimerization


  • Green Fluorescent Proteins

Associated data

  • PDB/4W69
  • PDB/4W6A
  • PDB/4W6B
  • PDB/4W6BS
  • PDB/4W6C
  • PDB/4W6D
  • PDB/4W6F
  • PDB/4W6G
  • PDB/4W6H
  • PDB/4W6I
  • PDB/4W6J
  • PDB/4W6K
  • PDB/4W6L
  • PDB/4W6M
  • PDB/4W6N
  • PDB/4W6O
  • PDB/4W6P
  • PDB/4W6R
  • PDB/4W6S
  • PDB/4W6T
  • PDB/4W6U
  • PDB/4W72
  • PDB/4W73
  • PDB/4W74
  • PDB/4W75
  • PDB/4W76
  • PDB/4W77
  • PDB/4W7A
  • PDB/4W7C
  • PDB/4W7D
  • PDB/4W7E
  • PDB/4W7F
  • PDB/4W7R
  • PDB/4W7X