Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis

Tamantha K. Littlejohn; Osamu Takikawa; Joanne F. Jamie; Mark J. Walker; Roger J. W. Truscott

doi:10.1016/S0531-5131(02)00312-6

Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis

Tamantha K. Littlejohn, Osamu Takikawa, Joanne F. Jamie, Mark J. Walker, Roger J. W. Truscott

Research output: Chapter in Book/Report/Conference proceeding › Conference proceeding contribution › peer-review

Abstract

Indoleamine 2,3-dioxygenase (IDO), the first enzyme of the kynurenine pathway of tryptophan metabolism, has been implicated in numerous disease states. Site-directed mutagenesis was undertaken using the expression plasmid pQE9-IDO, to incorporate a stop codon at lysine 389. This produced a C-terminally truncated protein, similar to that previously reported as a minor product during purification of the recombinant protein. Initial studies here, show that the Lys389 mutant retains enzymatic activity. Purification of this truncated protein, which lacks a presumably mobile terminal region, may increase the likelihood of crystallisation of human IDO.

Original language	English
Title of host publication	Oxygen and life
Subtitle of host publication	Oxygenases, oxidase, and lipid mediators
Editors	Y. Ishimura, M. Nozaki, S. Yamamoto, T. Shimizu, S. Narumiya, F. Mitani
Place of Publication	London
Publisher	Elsevier
Pages	157-160
Number of pages	4
Volume	1233
ISBN (Print)	0444508724, 9780444508720
DOIs	https://doi.org/10.1016/S0531-5131(02)00312-6
Publication status	Published - Nov 2002
Event	3rd International Conference on Oxygen and Life - KYOTO, Japan Duration: 26 Nov 2000 → 29 Nov 2000

Publication series

Name	INTERNATIONAL CONGRESS SERIES
Publisher	ELSEVIER SCIENCE BV
Volume	1233
ISSN (Print)	0531-5131

Conference

Conference	3rd International Conference on Oxygen and Life
Country/Territory	Japan
City	KYOTO
Period	26/11/00 → 29/11/00

Keywords

hexahistidyl tag
indoleamine 2,3-dioxygenase
site-directed mutagenesis

Access to Document

10.1016/S0531-5131(02)00312-6

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Littlejohn, T. K., Takikawa, O., Jamie, J. F., Walker, M. J., & Truscott, R. J. W. (2002). Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis. In Y. Ishimura, M. Nozaki, S. Yamamoto, T. Shimizu, S. Narumiya, & F. Mitani (Eds.), Oxygen and life: Oxygenases, oxidase, and lipid mediators (Vol. 1233, pp. 157-160). (INTERNATIONAL CONGRESS SERIES; Vol. 1233). Elsevier. https://doi.org/10.1016/S0531-5131(02)00312-6

Littlejohn, Tamantha K. ; Takikawa, Osamu ; Jamie, Joanne F. et al. / Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis. Oxygen and life: Oxygenases, oxidase, and lipid mediators. editor / Y. Ishimura ; M. Nozaki ; S. Yamamoto ; T. Shimizu ; S. Narumiya ; F. Mitani. Vol. 1233 London : Elsevier, 2002. pp. 157-160 (INTERNATIONAL CONGRESS SERIES).

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title = "Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis",

abstract = "Indoleamine 2,3-dioxygenase (IDO), the first enzyme of the kynurenine pathway of tryptophan metabolism, has been implicated in numerous disease states. Site-directed mutagenesis was undertaken using the expression plasmid pQE9-IDO, to incorporate a stop codon at lysine 389. This produced a C-terminally truncated protein, similar to that previously reported as a minor product during purification of the recombinant protein. Initial studies here, show that the Lys389 mutant retains enzymatic activity. Purification of this truncated protein, which lacks a presumably mobile terminal region, may increase the likelihood of crystallisation of human IDO.",

keywords = "hexahistidyl tag, indoleamine 2,3-dioxygenase, site-directed mutagenesis",

author = "Littlejohn, {Tamantha K.} and Osamu Takikawa and Jamie, {Joanne F.} and Walker, {Mark J.} and Truscott, {Roger J. W.}",

year = "2002",

month = nov,

doi = "10.1016/S0531-5131(02)00312-6",

language = "English",

isbn = "0444508724",

volume = "1233",

series = "INTERNATIONAL CONGRESS SERIES",

publisher = "Elsevier",

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editor = "Y. Ishimura and M. Nozaki and S. Yamamoto and T. Shimizu and S. Narumiya and F. Mitani",

booktitle = "Oxygen and life",

address = "Netherlands",

note = "3rd International Conference on Oxygen and Life ; Conference date: 26-11-2000 Through 29-11-2000",

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Littlejohn, TK, Takikawa, O, Jamie, JF, Walker, MJ & Truscott, RJW 2002, Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis. in Y Ishimura, M Nozaki, S Yamamoto, T Shimizu, S Narumiya & F Mitani (eds), Oxygen and life: Oxygenases, oxidase, and lipid mediators. vol. 1233, INTERNATIONAL CONGRESS SERIES, vol. 1233, Elsevier, London, pp. 157-160, 3rd International Conference on Oxygen and Life, KYOTO, Japan, 26/11/00. https://doi.org/10.1016/S0531-5131(02)00312-6

Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis. / Littlejohn, Tamantha K.; Takikawa, Osamu; Jamie, Joanne F. et al.
Oxygen and life: Oxygenases, oxidase, and lipid mediators. ed. / Y. Ishimura; M. Nozaki; S. Yamamoto; T. Shimizu; S. Narumiya; F. Mitani. Vol. 1233 London: Elsevier, 2002. p. 157-160 (INTERNATIONAL CONGRESS SERIES; Vol. 1233).

Research output: Chapter in Book/Report/Conference proceeding › Conference proceeding contribution › peer-review

TY - GEN

T1 - Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis

AU - Littlejohn, Tamantha K.

AU - Takikawa, Osamu

AU - Jamie, Joanne F.

AU - Walker, Mark J.

AU - Truscott, Roger J. W.

PY - 2002/11

Y1 - 2002/11

N2 - Indoleamine 2,3-dioxygenase (IDO), the first enzyme of the kynurenine pathway of tryptophan metabolism, has been implicated in numerous disease states. Site-directed mutagenesis was undertaken using the expression plasmid pQE9-IDO, to incorporate a stop codon at lysine 389. This produced a C-terminally truncated protein, similar to that previously reported as a minor product during purification of the recombinant protein. Initial studies here, show that the Lys389 mutant retains enzymatic activity. Purification of this truncated protein, which lacks a presumably mobile terminal region, may increase the likelihood of crystallisation of human IDO.

AB - Indoleamine 2,3-dioxygenase (IDO), the first enzyme of the kynurenine pathway of tryptophan metabolism, has been implicated in numerous disease states. Site-directed mutagenesis was undertaken using the expression plasmid pQE9-IDO, to incorporate a stop codon at lysine 389. This produced a C-terminally truncated protein, similar to that previously reported as a minor product during purification of the recombinant protein. Initial studies here, show that the Lys389 mutant retains enzymatic activity. Purification of this truncated protein, which lacks a presumably mobile terminal region, may increase the likelihood of crystallisation of human IDO.

KW - hexahistidyl tag

KW - indoleamine 2,3-dioxygenase

KW - site-directed mutagenesis

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U2 - 10.1016/S0531-5131(02)00312-6

DO - 10.1016/S0531-5131(02)00312-6

M3 - Conference proceeding contribution

SN - 0444508724

SN - 9780444508720

VL - 1233

T3 - INTERNATIONAL CONGRESS SERIES

SP - 157

EP - 160

BT - Oxygen and life

A2 - Ishimura, Y.

A2 - Nozaki, M.

A2 - Yamamoto, S.

A2 - Shimizu, T.

A2 - Narumiya, S.

A2 - Mitani, F.

PB - Elsevier

CY - London

T2 - 3rd International Conference on Oxygen and Life

Y2 - 26 November 2000 through 29 November 2000

ER -

Littlejohn TK, Takikawa O, Jamie JF, Walker MJ, Truscott RJW. Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis. In Ishimura Y, Nozaki M, Yamamoto S, Shimizu T, Narumiya S, Mitani F, editors, Oxygen and life: Oxygenases, oxidase, and lipid mediators. Vol. 1233. London: Elsevier. 2002. p. 157-160. (INTERNATIONAL CONGRESS SERIES). doi: 10.1016/S0531-5131(02)00312-6

Production of truncated enzymically active human indoleamine 2,3-dioxygenase using site-directed mutagenesis

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