Luciola cruciata luciferase: Difference between revisions
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2D1R is a 1 chain structure of sequence from [http://en.wikipedia.org/wiki/Luciola_cruciata Luciola cruciata]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2D1R OCA]. | 2D1R is a 1 chain structure of sequence from [http://en.wikipedia.org/wiki/Luciola_cruciata Luciola cruciata]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2D1R OCA]. | ||
The protein is shown here bound with two products, adenosine 5'-phosphate (AMP) and oxyluciferine. | The protein is shown here bound with two products, adenosine 5'-phosphate (AMP) and oxyluciferine. | ||
It weights 62 kDa and is constituted of two main domains, a large N-terminal domain and a small C-terminal domain, separated by a cleft and linked by a hinge. <scene name='Luciola_cruciata_luciferase/Nter_domain/2'>The N-ter domain</scene> (residues 1-436) is made of a β-barrel and two β-sheets, flanked by α-helices, thus forming | It weights 62 kDa and is constituted of two main domains, a large N-terminal domain and a small C-terminal domain, separated by a cleft and linked by a hinge. <scene name='Luciola_cruciata_luciferase/Nter_domain/2'>The N-ter domain</scene> (residues 1-436) is made of a β-barrel and two β-sheets, flanked by α-helices, thus forming | ||
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There are two conformational states, one when the enzyme is substrate-free, and the other one product-bound (whose structure is represented on this page). The transition is achieved by a 90° counterclockwise rotation of the <scene name='Luciola_cruciata_luciferase/Cter_relative_nter/1'>C-domain relative to the N-domain</scene><ref name="ane">PMID:17513367</ref> | There are two conformational states, one when the enzyme is substrate-free, and the other one product-bound (whose structure is represented on this page). The transition is achieved by a 90° counterclockwise rotation of the <scene name='Luciola_cruciata_luciferase/Cter_relative_nter/1'>C-domain relative to the N-domain</scene><ref name="ane">PMID:17513367</ref> | ||
===Sequence similarities=== | ===Sequence similarities=== | ||
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But the firefly luciferase also have caracteristic sequences found in no other nucleotide-binding proteins, such as a motif responsible for the binding of ATP. | But the firefly luciferase also have caracteristic sequences found in no other nucleotide-binding proteins, such as a motif responsible for the binding of ATP. | ||
===Spectral difference with mutated luciferases=== | ===Spectral difference with mutated luciferases=== | ||
The colour changes from the classical yellow-green colour to red with the substitution of a single amino acid, <scene name='Sandbox2442/Residue_ser_286/1'>Ser286</scene> to Asn, in the S286N mutant. The active site (and the residue Ile288) is less potent to effectue it's conformational change to the closed state, which was providing an extremely hydrophobic environment. This is then allowing an energy loss, and the product will emit lower energy light, with a wavelength moved to the red. | The colour changes from the classical yellow-green colour to red with the substitution of a single amino acid, <scene name='Sandbox2442/Residue_ser_286/1'>Ser286</scene> to Asn, in the S286N mutant. The active site (and the residue Ile288) is less potent to effectue it's conformational change to the closed state, which was providing an extremely hydrophobic environment. This is then allowing an energy loss, and the product will emit lower energy light, with a wavelength moved to the red. | ||
Further mutations proved that the <scene name='Sandbox2442/Ile_288/1'>amino acid residue at position 288</scene> is influencing the wavelength of the light emitted.<ref>PMID:16541080</ref> | Further mutations proved that the <scene name='Sandbox2442/Ile_288/1'>amino acid residue at position 288</scene> is influencing the wavelength of the light emitted.<ref>PMID:16541080</ref> | ||
=== High-energy intermediate analogue=== | === High-energy intermediate analogue=== | ||
<scene name='Luciferase/2d1s/2'>Luciferase</scene> ([[2d1s]]). | |||
Generally, firefly luciferases have some similarities with Acyl-CoA ligases and some peptide synthetases despite having different cellular effects. In fixing the structure of L. cruciata luciferase, the analog of a potent aminoacyl-tRNA synthetases (DLSA) was successfuly utilized to represent a stable oxyluciferin intermediate.<ref name="structure">PMID:16541080 </ref>.<br> | Generally, firefly luciferases have some similarities with Acyl-CoA ligases and some peptide synthetases despite having different cellular effects. In fixing the structure of L. cruciata luciferase, the analog of a potent aminoacyl-tRNA synthetases (DLSA) was successfuly utilized to represent a stable oxyluciferin intermediate.<ref name="structure">PMID:16541080 </ref>.<br> | ||
{{Link Toggle FancyCartoonHighQualityView}} on 2d1s (see right image).<br> | {{Link Toggle FancyCartoonHighQualityView}} on 2d1s (see right image).<br> | ||
