Crystal structures of green fluorescent protein with the unnatural amino acid 4-nitro-L -phenylalanine

Maurici, Nicole; Savidge, Nicole; Lee, Byung Uk; Brewer, Scott H.; Phillips-Piro, Christine M.

doi:10.1107/S2053230X1801169X

Title: Crystal structures of green fluorescent protein with the unnatural amino acid 4-nitro-L -phenylalanine

Journal Article · Wed Sep 19 00:00:00 EDT 2018 · Acta Crystallographica. Section F, Structural Biology Communications

DOI:https://doi.org/10.1107/S2053230X1801169X· OSTI ID:1483086

Maurici, Nicole; Savidge, Nicole; Lee, Byung Uk; Brewer, Scott H.;

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 (pNO₂F), at two unique sites in the protein have been determined. Amber codon-suppression methodology was used to site-specifically incorporate pNO₂F at a solvent-accessible (Asp133) and a partially buried (Asn149) site in sfGFP. The Asp133pNO₂F sfGFP construct crystallized with two molecules per asymmetric unit in space group P3₂21 and the crystal structure was refined to 2.05 Å resolution. Crystals of Asn149pNO₂F sfGFP contained one molecule of sfGFP per asymmetric unit in space group P4₁22 and the structure was refined to 1.60 Å resolution. The alignment of Asp133pNO₂F or Asn149pNO₂F 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 pNO₂F as an effective spectroscopic reporter of local protein structure and dynamics.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: OTHER

OSTI ID:: 1483086

Journal Information:: Acta Crystallographica. Section F, Structural Biology Communications, Vol. 74, Issue 10; ISSN 2053-230X

Publisher:: International Union of Crystallography

Country of Publication:: United States

Language:: ENGLISH

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