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 proceedingConference proceeding contributionpeer-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 languageEnglish
Title of host publicationOxygen and life
Subtitle of host publicationOxygenases, oxidase, and lipid mediators
EditorsY. Ishimura, M. Nozaki, S. Yamamoto, T. Shimizu, S. Narumiya, F. Mitani
Place of PublicationLondon
PublisherElsevier
Pages157-160
Number of pages4
Volume1233
ISBN (Print)0444508724, 9780444508720
DOIs
Publication statusPublished - Nov 2002
Event3rd International Conference on Oxygen and Life - KYOTO, Japan
Duration: 26 Nov 200029 Nov 2000

Publication series

NameINTERNATIONAL CONGRESS SERIES
PublisherELSEVIER SCIENCE BV
Volume1233
ISSN (Print)0531-5131

Conference

Conference3rd International Conference on Oxygen and Life
CountryJapan
CityKYOTO
Period26/11/0029/11/00

Keywords

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

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