 |
PDBsum entry 1oxe
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Luminescent protein
|
PDB id
|
|
|
|
1oxe
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Expansion of the genetic code enables design of a novel "gold" class of green fluorescent proteins.
|
|
|
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.
|
|
|
Ref.
|
|
J Mol Biol, 2003,
328,
1071-1081.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
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.
|
|
|
|
|
|
|
|
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.
|
|
|
|
|
|
|
