6DQ1

sfGFP N149 mutated to 4-nitro-L-phenylalanine

Summary for 6DQ1

• Entry DOI: 10.2210/pdb6dq1/pdb
• Related: 6DQ0
• Descriptor: sfGFP, SODIUM ION (3 entities in total)
• Functional Keywords: super folder gfp, 4-nitro-l-phenylalanine, unnatural amino acid, fluorescent protein
• Biological source: Aequorea victoria
• Total number of polymer chains: 1
• Total formula weight: 26999.39

Authors

Phillips-Piro, C.M.,Savidge, N.,Lee, B. (deposition date: 2018-06-10, release date: 2018-10-17, Last modification date: 2023-11-15)

Primary citation

Maurici, N.,Savidge, N.,Lee, B.U.,Brewer, S.H.,Phillips-Piro, C.M.
Crystal structures of green fluorescent protein with the unnatural amino acid 4-nitro-L-phenylalanine.
Acta Crystallogr F Struct Biol Commun, 74:650-655, 2018

PubMed Abstract: The X-ray crystal structures of two superfolder green fluorescent protein (sfGFP) constructs containing a genetically incorporated spectroscopic reporter unnatural amino acid, 4-nitro-L-phenylalanine (pNOF), at two unique sites in the protein have been determined. Amber codon-suppression methodology was used to site-specifically incorporate pNOF at a solvent-accessible (Asp133) and a partially buried (Asn149) site in sfGFP. The Asp133pNOF sfGFP construct crystallized with two molecules per asymmetric unit in space group P321 and the crystal structure was refined to 2.05 Å resolution. Crystals of Asn149pNOF sfGFP contained one molecule of sfGFP per asymmetric unit in space group P422 and the structure was refined to 1.60 Å resolution. The alignment of Asp133pNOF or Asn149pNOF sfGFP with wild-type sfGFP resulted in small root-mean-square deviations, illustrating that these residues do not significantly alter the protein structure and supporting the use of pNOF as an effective spectroscopic reporter of local protein structure and dynamics.

PubMed: 30279317
DOI: 10.1107/S2053230X1801169X

Experimental method

X-RAY DIFFRACTION (1.598 Å)