5DPI

sfGFP double mutant - 133/149 p-cyano-L-phenylalanine

Summary for 5DPI

• Entry DOI: 10.2210/pdb5dpi/pdb
• Related: 5DPG 5DPH 5DPJ
• Descriptor: Green fluorescent protein (2 entities in total)
• Functional Keywords: gfp, unnatural amino acid, cyanophenylalanine, fluorescent protein
• Biological source: Aequorea victoria (Jellyfish)
• Total number of polymer chains: 6
• Total formula weight: 162081.02

Authors

Dippel, A.B.,Olenginski, G.M.,Maurici, N.,Liskov, M.T.,Brewer, S.H.,Phillips-Piro, C.M. (deposition date: 2015-09-12, release date: 2016-01-13, Last modification date: 2019-11-27)

Primary citation

Dippel, A.B.,Olenginski, G.M.,Maurici, N.,Liskov, M.T.,Brewer, S.H.,Phillips-Piro, C.M.
Probing the effectiveness of spectroscopic reporter unnatural amino acids: a structural study.
Acta Crystallogr D Struct Biol, 72:121-130, 2016

PubMed Abstract: The X-ray crystal structures of superfolder green fluorescent protein (sfGFP) containing the spectroscopic reporter unnatural amino acids (UAAs) 4-cyano-L-phenylalanine (pCNF) or 4-ethynyl-L-phenylalanine (pCCF) at two unique sites in the protein have been determined. These UAAs were genetically incorporated into sfGFP in a solvent-exposed loop region and/or a partially buried site on the β-barrel of the protein. The crystal structures containing the UAAs at these two sites permit the structural implications of UAA incorporation for the native protein structure to be assessed with high resolution and permit a direct correlation between the structure and spectroscopic data to be made. The structural implications were quantified by comparing the root-mean-square deviation (r.m.s.d.) between the crystal structure of wild-type sfGFP and the protein constructs containing either pCNF or pCCF in the local environment around the UAAs and in the overall protein structure. The results suggest that the selective placement of these spectroscopic reporter UAAs permits local protein environments to be studied in a relatively nonperturbative fashion with site-specificity.

PubMed: 26894540
DOI: 10.1107/S2059798315022858

Experimental method

X-RAY DIFFRACTION (2.54 Å)