4LW5

Crystal structure of all-trans green fluorescent protein

Summary for 4LW5

• Entry DOI: 10.2210/pdb4lw5/pdb
• Descriptor: Green fluorescent protein (2 entities in total)
• Functional Keywords: 11-stranded beta barrel, fluorescent protein
• Biological source: Aequorea victoria (Jellyfish)
• Total number of polymer chains: 5
• Total formula weight: 134701.73

Authors

Rosenman, D.J.,Huang, Y.-M.,Xia, K.,Vanroey, P.,Colon, W.,Bystroff, C. (deposition date: 2013-07-26, release date: 2014-02-05, Last modification date: 2024-11-20)

Primary citation

Rosenman, D.J.,Huang, Y.M.,Xia, K.,Fraser, K.,Jones, V.E.,Lamberson, C.M.,Van Roey, P.,Colon, W.,Bystroff, C.
Green-lighting green fluorescent protein: Faster and more efficient folding by eliminating a cis-trans peptide isomerization event.
Protein Sci., 23:400-410, 2014

PubMed Abstract: Wild-type green fluorescent protein (GFP) folds on a time scale of minutes. The slow step in folding is a cis-trans peptide bond isomerization. The only conserved cis-peptide bond in the native GFP structure, at P89, was remodeled by the insertion of two residues, followed by iterative energy minimization and side chain design. The engineered GFP was synthesized and found to fold faster and more efficiently than its template protein, recovering 50% more of its fluorescence upon refolding. The slow phase of folding is faster and smaller in amplitude, and hysteresis in refolding has been eliminated. The elimination of a previously reported kinetically trapped state in refolding suggests that X-P89 is trans in the trapped state. A 2.55 Å resolution crystal structure revealed that the new variant contains only trans-peptide bonds, as designed. This is the first instance of a computationally remodeled fluorescent protein that folds faster and more efficiently than wild type.

PubMed: 24408076
DOI: 10.1002/pro.2421

Experimental method

X-RAY DIFFRACTION (2.55 Å)