PDBsum entry 1oxe

Luminescent protein

PDB id: 1oxe

Contents

Protein chain

227 a.a. *

Waters ×166

* Residue conservation analysis

PDB id:

1oxe

Name:

Luminescent protein

Title:

Expansion of the genetic code enables design of a novel "gold" class of green fluorescent proteins

Structure:

Cyan fluorescent protein cfp. Chain: a. Engineered: yes. Mutation: yes

Source:

Cfp marker plasmid pwm1009. Organism_taxid: 141850. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.15Å R-factor: 0.223 R-free: 0.236

Authors:

J.Hyun Bae,M.Rubini,G.Jung,G.Wiegand,M.H.Seifert,M.K.Azim,J.S.Kim, A.Zumbusch,T.A.Holak,L.Moroder,R.Huber,N.Budisa

Key ref:

J.Hyun Bae et al. (2003). Expansion of the genetic code enables design of a novel "gold" class of green fluorescent proteins. J Mol Biol, 328, 1071-1081. PubMed id: 12729742 DOI: 10.1016/S0022-2836(03)00364-4

Date:

02-Apr-03 Release date: 02-Dec-03

Protein chain

No UniProt id for this chain

Struc: 227 a.a.

DOI no: 10.1016/S0022-2836(03)00364-4

J Mol Biol 328:1071-1081 (2003)

PubMed id: 12729742

Expansion of the genetic code enables design of a novel "gold" class of green fluorescent proteins.

J.Hyun Bae, M.Rubini, G.Jung, G.Wiegand, M.H.Seifert, M.K.Azim, J.S.Kim, A.Zumbusch, T.A.Holak, L.Moroder, R.Huber, N.Budisa.

ABSTRACT

Much effort has been dedicated to the design of significantly red shifted variants of the green fluorescent protein (GFP) from Aequoria victora (av). These approaches have been based on classical engineering with the 20 canonical amino acids. We report here an expansion of these efforts by incorporation of an amino substituted variant of tryptophan into the "cyan" GFP mutant, which turned it into a "gold" variant. This variant possesses a red shift in emission unprecedented for any avFP, similar to "red" FPs, but with enhanced stability and a very low aggregation tendency. An increasing number of non-natural amino acids are available for chromophore redesign (by engineering of the genetic code) and enable new general strategies to generate novel classes of tailor-made GFP proteins.

Selected figure(s)

Figure 2.
Figure 2. Spectral properties of ECFP and EGFP with substituted Trp residues. Spectra were measured at 20 °C in aqueous buffered solution. In ECFP Trp57 and chromophore Trp66 replacement with (4-NH[2])-Trp yielded the novel protein variant GdFP with golden colour. Molar extinction is expressed in M -1 cm -1.

Figure 5.
Figure 5. (i) Overall structure of GFPs. C^a traces are coloured blue (ECFP), yellow (GdFP) and green (EGFP), drawn by MolMol.[29.] The chromophore and residues Tyr145 and His148 are represented as sticks in the same colours with the exception of ECFP where blue (conformation A' of ECFP), and red (conformation B' of ECFP) are used. (ii) Schematic representation of ECFP (major configuration A') and GdFP protein matrix-chromophore interactions drawn by LIGPLOT. [30.] In the minor conformation B', Ile146 makes a hydrophobic contact with the chromopore while Tyr145 does not (see Supplementary Material). (iii) Stereo-view of the chromophores in EGFP (green) ECFP (blue) and GdFP (yellow) and their environments. Tyr145 and His148 of the ECFP minor-form are shown in red. Note a slight shift of the amino indole moiety of the GdFP chromophore toward the Phe165, a property unique among other GFPs.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2003, 328, 1071-1081) copyright 2003.

Figures were selected by an automated process.