Abstract
Methods are presented for the use of the coral 8R-lipoxygenase from the Caribbean sea whip coral Plexaura homomalla as a model enzyme for structural studies of animal lipoxygenases. The 8R-lipoxygenase is remarkably stable and can be stored at 4°C for 3 months with virtually no loss of activity. In addition, an engineered "pseudo wild-type" enzyme is soluble in the absence of detergents, which helps facilitate the preparation of enzyme:substrate complexes.
Keywords:
Anaerobic; Crystallization; Enzyme–substrate complex; Expression; Lipoxygenase; Purification; Structural biology.
© 2018 Elsevier Inc. All rights reserved.
MeSH terms
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Animals
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Anthozoa / metabolism*
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Arachidonate Lipoxygenases / chemistry
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Arachidonate Lipoxygenases / genetics
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Arachidonate Lipoxygenases / isolation & purification*
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Arachidonate Lipoxygenases / metabolism
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Arachidonic Acid / chemistry
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Arachidonic Acid / metabolism*
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Binding Sites / genetics
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Crystallography, X-Ray / methods
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Enzyme Assays / methods*
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Escherichia coli / metabolism
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Metabolic Engineering / methods
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Protein Domains / genetics*
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Substrate Specificity / genetics
Substances
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Recombinant Proteins
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Arachidonic Acid
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Arachidonate Lipoxygenases
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arachidonate 8-lipoxygenase