Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()

Kynurenine 3-monooxygenase (KMO) is an enzyme central to the kynurenine pathway of tryptophan metabolism. KMO has been implicated as a therapeutic target in several disease states, including Huntington’s disease. Recombinant human KMO protein production is challenging due to the presence of transmem...

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Autores principales: Wilson, K., Mole, D.J., Binnie, M., Homer, N.Z.M., Zheng, X., Yard, B.A., Iredale, J.P., Auer, M., Webster, S.P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2014
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3969302/
https://www.ncbi.nlm.nih.gov/pubmed/24316190
http://dx.doi.org/10.1016/j.pep.2013.11.015
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author Wilson, K.
Mole, D.J.
Binnie, M.
Homer, N.Z.M.
Zheng, X.
Yard, B.A.
Iredale, J.P.
Auer, M.
Webster, S.P.
author_facet Wilson, K.
Mole, D.J.
Binnie, M.
Homer, N.Z.M.
Zheng, X.
Yard, B.A.
Iredale, J.P.
Auer, M.
Webster, S.P.
author_sort Wilson, K.
collection PubMed
description Kynurenine 3-monooxygenase (KMO) is an enzyme central to the kynurenine pathway of tryptophan metabolism. KMO has been implicated as a therapeutic target in several disease states, including Huntington’s disease. Recombinant human KMO protein production is challenging due to the presence of transmembrane domains, which localise KMO to the outer mitochondrial membrane and render KMO insoluble in many in vitro expression systems. Efficient bacterial expression of human KMO would accelerate drug development of KMO inhibitors but until now this has not been achieved. Here we report the first successful bacterial (Escherichia coli) expression of active FLAG™-tagged human KMO enzyme expressed in the soluble fraction and progress towards its purification.
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spelling pubmed-39693022014-03-31 Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification() Wilson, K. Mole, D.J. Binnie, M. Homer, N.Z.M. Zheng, X. Yard, B.A. Iredale, J.P. Auer, M. Webster, S.P. Protein Expr Purif Article Kynurenine 3-monooxygenase (KMO) is an enzyme central to the kynurenine pathway of tryptophan metabolism. KMO has been implicated as a therapeutic target in several disease states, including Huntington’s disease. Recombinant human KMO protein production is challenging due to the presence of transmembrane domains, which localise KMO to the outer mitochondrial membrane and render KMO insoluble in many in vitro expression systems. Efficient bacterial expression of human KMO would accelerate drug development of KMO inhibitors but until now this has not been achieved. Here we report the first successful bacterial (Escherichia coli) expression of active FLAG™-tagged human KMO enzyme expressed in the soluble fraction and progress towards its purification. Academic Press 2014-03 /pmc/articles/PMC3969302/ /pubmed/24316190 http://dx.doi.org/10.1016/j.pep.2013.11.015 Text en © 2013 The Authors http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Article
Wilson, K.
Mole, D.J.
Binnie, M.
Homer, N.Z.M.
Zheng, X.
Yard, B.A.
Iredale, J.P.
Auer, M.
Webster, S.P.
Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()
title Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()
title_full Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()
title_fullStr Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()
title_full_unstemmed Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()
title_short Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification()
title_sort bacterial expression of human kynurenine 3-monooxygenase: solubility, activity, purification()
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3969302/
https://www.ncbi.nlm.nih.gov/pubmed/24316190
http://dx.doi.org/10.1016/j.pep.2013.11.015
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