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PDBsum entry 2d1t
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Oxidoreductase
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PDB id
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2d1t
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References listed in PDB file
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Key reference
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Title
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Structural basis for the spectral difference in luciferase bioluminescence.
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Authors
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T.Nakatsu,
S.Ichiyama,
J.Hiratake,
A.Saldanha,
N.Kobashi,
K.Sakata,
H.Kato.
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Ref.
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Nature, 2006,
440,
372-376.
[DOI no: ]
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PubMed id
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Abstract
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Fireflies communicate with each other by emitting yellow-green to yellow-orange
brilliant light. The bioluminescence reaction, which uses luciferin, Mg-ATP and
molecular oxygen to yield an electronically excited oxyluciferin species, is
carried out by the enzyme luciferase. Visible light is emitted during relaxation
of excited oxyluciferin to its ground state. The high quantum yield of the
luciferin/luciferase reaction and the change in bioluminescence colour caused by
subtle structural differences in luciferase have attracted much research
interest. In fact, a single amino acid substitution in luciferase changes the
emission colour from yellow-green to red. Although the crystal structure of
luciferase from the North American firefly (Photinus pyralis) has been
described, the detailed mechanism for the bioluminescence colour change is still
unclear. Here we report the crystal structures of wild-type and red mutant
(S286N) luciferases from the Japanese Genji-botaru (Luciola cruciata) in complex
with a high-energy intermediate analogue,
5'-O-[N-(dehydroluciferyl)-sulfamoyl]adenosine (DLSA). Comparing these
structures to those of the wild-type luciferase complexed with AMP plus
oxyluciferin (products) reveals a significant conformational change in the
wild-type enzyme but not in the red mutant. This conformational change involves
movement of the hydrophobic side chain of Ile 288 towards the benzothiazole ring
of DLSA. Our results indicate that the degree of molecular rigidity of the
excited state of oxyluciferin, which is controlled by a transient movement of
Ile 288, determines the colour of bioluminescence during the emission reaction.
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Figure 1.
Figure 1: The bioluminescence reaction catalysed by luciferase.
Figure 1 : The bioluminescence reaction catalysed by
luciferase. Unfortunately we are unable to provide accessible
alternative text for this. If you require assistance to access
this image, or to obtain a text description, please contact
npg@nature.com-
a, A two-step reaction mechanism via the luciferyl  AMP
intermediate. b, Structure of DLSA, a luciferyl AMP
intermediate analogue.
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Figure 4.
Figure 4: Bioluminescence colour of wild-type and three mutant
forms (I288V, I288A and S286N) of Lcr luciferases. a,
Photographs of bioluminescence by a luciferase-catalysed
reaction. The reaction condition was 0.1 mM luciferin, 2 mM ATP,
6 mM MgSO[4], 25 mM HEPES (pH 7.8) and 0.1 mg ml^-1 luciferase
in 0.5 ml solution. b, Emission spectra of the four types of
Luciola cruciata luciferases.
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nature
(2006,
440,
372-376)
copyright 2006.
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