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PDBsum entry 4j2p
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Luminescent protein
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PDB id
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4j2p
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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 and biochemical properties of luxf from photobacterium leiognathi.
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Authors
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T.Bergner,
C.R.Tabib,
A.Winkler,
S.Stipsits,
H.Kayer,
J.Lee,
J.P.Malthouse,
S.Mayhew,
F.Müller,
K.Gruber,
P.Macheroux.
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Ref.
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Biochim Biophys Acta, 2015,
1854,
1466-1475.
[DOI no: ]
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PubMed id
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Abstract
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The lux-operon of bioluminescent bacteria contains the genes coding for the
enzymes required for light emission. Some species of Photobacteria feature an
additional gene, luxF, which shows similarity to luxA and luxB, the genes
encoding the heterodimeric luciferase. Isolated dimeric LuxF binds four
molecules of an unusually derivatized flavin, i.e., 6-(3'-(R)-myristyl)-FMN
(myrFMN). In the present study we have heterologously expressed LuxF in
Escherichia coli BL21 in order to advance our understanding of the protein's
binding properties and its role in photobacterial bioluminescence. Structure
determination by X-ray crystallography confirmed that apo-LuxF possesses four
preorganized binding sites, which are further optimized by adjusting the
orientation of amino acid side chains. To investigate the binding properties of
recombinant LuxF we have isolated myrFMN from Photobacterium leiognathi S1. We
found that LuxF binds myrFMN tightly with a dissociation constant of 80±20 nM
demonstrating that the purified apo-form of LuxF is fully competent in myrFMN
binding. In contrast to LuxF, binding of myrFMN to luciferase is much weaker
(Kd=4.0±0.4 μM) enabling LuxF to prevent inhibition of the enzyme by
scavenging myrFMN. Moreover, we have used apo-LuxF to demonstrate that myrFMN
occurs in all Photobacteria tested, irrespective of the presence of luxF
indicating that LuxF is not required for myrFMN biosynthesis.
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