Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production

Bellucci, Michele; Pompa, Andrea; De Marcos Lousa, Carine; Panfili, Eleonora; Orecchini, Elena; Maricchiolo, Elisa; Fraternale, Daniele; Orabona, Ciriana; De Marchis, Francesca; Pallotta, Maria Teresa

doi:10.3390/ijms22105102

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Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production

Michele Bellucci

^1,†

Andrea Pompa

^1,2,†

Carine De Marcos Lousa

^3,4

⁵,

⁵

²,

⁵

^1,*

and

Maria Teresa Pallotta

^5,*

Institute of Biosciences and Bioresources, National Research Council of Italy, 06128 Perugia, Italy

Department of Biomolecular Sciences, University Carlo Bo, 61029 Urbino, Italy

Centre for Biomedical Sciences, School of Clinical and Applied Sciences, Leeds Beckett University, Leeds LS13HE, UK

⁴

Centre for Plant Sciences, Faculty of Biological Sciences, University of Leeds, Leeds LS29JT, UK

⁵

Department of Medicine and Surgery, University of Perugia, 06128 Perugia, Italy

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Academic Editor: Pavel Soucek

Int. J. Mol. Sci. 2021, 22(10), 5102; https://doi.org/10.3390/ijms22105102

Received: 24 March 2021 / Revised: 21 April 2021 / Accepted: 8 May 2021 / Published: 12 May 2021

(This article belongs to the Special Issue Decoding the Complex Crossroad of Tryptophan Metabolic Pathways)

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Abstract

Genetic engineering of plants has turned out to be an attractive approach to produce various secondary metabolites. Here, we attempted to produce kynurenine, a health-promoting metabolite, in plants of Nicotiana tabacum (tobacco) transformed by Agrobacterium tumefaciens with the gene, coding for human indoleamine 2,3-dioxygenase 1 (IDO1), an enzyme responsible for the kynurenine production because of tryptophan degradation. The presence of IDO1 gene in transgenic plants was confirmed by PCR, but the protein failed to be detected. To confer higher stability to the heterologous human IDO1 protein and to provide a more sensitive method to detect the protein of interest, we cloned a gene construct coding for IDO1-GFP. Analysis of transiently transfected tobacco protoplasts demonstrated that the IDO1-GFP gene led to the expression of a detectable protein and to the production of kynurenine in the protoplast medium. Interestingly, the intracellular localisation of human IDO1 in plant cells is similar to that found in mammal cells, mainly in cytosol, but in early endosomes as well. To the best of our knowledge, this is the first report on the expression of human IDO1 enzyme capable of secreting kynurenines in plant cells. View Full-Text

Keywords: protoplasts; kynurenine; genetic transformation

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Bellucci, M.; Pompa, A.; De Marcos Lousa, C.; Panfili, E.; Orecchini, E.; Maricchiolo, E.; Fraternale, D.; Orabona, C.; De Marchis, F.; Pallotta, M.T. Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production. Int. J. Mol. Sci. 2021, 22, 5102. https://doi.org/10.3390/ijms22105102

AMA Style

Bellucci M, Pompa A, De Marcos Lousa C, Panfili E, Orecchini E, Maricchiolo E, Fraternale D, Orabona C, De Marchis F, Pallotta MT. Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production. International Journal of Molecular Sciences. 2021; 22(10):5102. https://doi.org/10.3390/ijms22105102

Chicago/Turabian Style

Bellucci, Michele, Andrea Pompa, Carine De Marcos Lousa, Eleonora Panfili, Elena Orecchini, Elisa Maricchiolo, Daniele Fraternale, Ciriana Orabona, Francesca De Marchis, and Maria T. Pallotta. 2021. "Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production" International Journal of Molecular Sciences 22, no. 10: 5102. https://doi.org/10.3390/ijms22105102

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Human Indoleamine 2,3-dioxygenase 1 (IDO1) Expressed in Plant Cells Induces Kynurenine Production

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