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

K Wilson; D J Mole; M Binnie; N Z M Homer; X Zheng; B A Yard; J P Iredale; M Auer; S P Webster

doi:10.1016/j.pep.2013.11.015

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

K Wilson, D J Mole, M Binnie, N Z M Homer, X Zheng, B A Yard, J P Iredale, M Auer, S P Webster

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Pages (from-to)	96-103
Number of pages	8
Journal	Protein Expression and Purification
Volume	95
DOIs	https://doi.org/10.1016/j.pep.2013.11.015
Publication status	Published - Mar 2014

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10.1016/j.pep.2013.11.015

Bacterial expression of human kynurenine 3-monooxygenase Solubility, activity, purification
Open Access funded by Medical Research Council License: http://creativecommons.org/licenses/by/3.0/
Final published version, 1.27 MBLicence: Creative Commons: Attribution (CC-BY)

http://www.sciencedirect.com/science/article/pii/S1046592813002593

PhD STUDENT - JOANNE SIMPSON - Supervisor MOHINI GRAY
Iredale, J.
MRC
1/09/12 → 31/08/16
Project: Research

Cite this

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title = "Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification",

abstract = "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{\texttrademark}-tagged human KMO enzyme expressed in the soluble fraction and progress towards its purification.",

author = "K Wilson and Mole, {D J} and M Binnie and Homer, {N Z M} and X Zheng and Yard, {B A} and Iredale, {J P} and M Auer and Webster, {S P}",

note = "This work was supported by the Medical Research Council.",

year = "2014",

month = mar,

doi = "10.1016/j.pep.2013.11.015",

language = "English",

volume = "95",

pages = "96--103",

journal = "Protein Expression and Purification",

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TY - JOUR

T1 - Bacterial expression of human kynurenine 3-monooxygenase

T2 - Solubility, activity, purification

AU - Wilson, K

AU - Mole, D J

AU - Binnie, M

AU - Homer, N Z M

AU - Zheng, X

AU - Yard, B A

AU - Iredale, J P

AU - Auer, M

AU - Webster, S P

N1 - This work was supported by the Medical Research Council.

PY - 2014/3

Y1 - 2014/3

N2 - 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.

AB - 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.

U2 - 10.1016/j.pep.2013.11.015

DO - 10.1016/j.pep.2013.11.015

M3 - Article

C2 - 24316190

SN - 1046-5928

VL - 95

SP - 96

EP - 103

JO - Protein Expression and Purification

JF - Protein Expression and Purification

ER -

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

Abstract

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PhD STUDENT - JOANNE SIMPSON - Supervisor MOHINI GRAY

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