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A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin

While heme synthesis requires the formation of a potentially lethal intermediate, protoporphyrin IX (PPIX), surprisingly little is known about the mechanism of its toxicity, aside from its phototoxicity. The cellular protein interactions of PPIX might provide insight into modulators of PPIX-induced...

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Autores principales: Lynch, John, Wang, Yao, Li, Yuxin, Kavdia, Kanisha, Fukuda, Yu, Ranjit, Sabina, Robinson, Camenzind G., Grace, Christy R., Xia, Youlin, Peng, Junmin, Schuetz, John D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10290680/
https://www.ncbi.nlm.nih.gov/pubmed/37355765
http://dx.doi.org/10.1038/s42003-023-05024-5
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author Lynch, John
Wang, Yao
Li, Yuxin
Kavdia, Kanisha
Fukuda, Yu
Ranjit, Sabina
Robinson, Camenzind G.
Grace, Christy R.
Xia, Youlin
Peng, Junmin
Schuetz, John D.
author_facet Lynch, John
Wang, Yao
Li, Yuxin
Kavdia, Kanisha
Fukuda, Yu
Ranjit, Sabina
Robinson, Camenzind G.
Grace, Christy R.
Xia, Youlin
Peng, Junmin
Schuetz, John D.
author_sort Lynch, John
collection PubMed
description While heme synthesis requires the formation of a potentially lethal intermediate, protoporphyrin IX (PPIX), surprisingly little is known about the mechanism of its toxicity, aside from its phototoxicity. The cellular protein interactions of PPIX might provide insight into modulators of PPIX-induced cell death. Here we report the development of PPB, a biotin-conjugated, PPIX-probe that captures proteins capable of interacting with PPIX. Quantitative proteomics in a diverse panel of mammalian cell lines reveal a high degree of concordance for PPB-interacting proteins identified for each cell line. Most differences are quantitative, despite marked differences in PPIX formation and sensitivity. Pathway and quantitative difference analysis indicate that iron and heme metabolism proteins are prominent among PPB-bound proteins in fibroblasts, which undergo PPIX-mediated death determined to occur through ferroptosis. PPB proteomic data (available at PRIDE ProteomeXchange # PXD042631) reveal that redox proteins from PRDX family of glutathione peroxidases interact with PPIX. Targeted gene knockdown of the mitochondrial PRDX3, but not PRDX1 or 2, enhance PPIX-induced death in fibroblasts, an effect blocked by the radical-trapping antioxidant, ferrostatin-1. Increased PPIX formation and death was also observed in a T-lymphoblastoid ferrochelatase-deficient leukemia cell line, suggesting that PPIX elevation might serve as a potential strategy for killing certain leukemias.
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spelling pubmed-102906802023-06-26 A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin Lynch, John Wang, Yao Li, Yuxin Kavdia, Kanisha Fukuda, Yu Ranjit, Sabina Robinson, Camenzind G. Grace, Christy R. Xia, Youlin Peng, Junmin Schuetz, John D. Commun Biol Article While heme synthesis requires the formation of a potentially lethal intermediate, protoporphyrin IX (PPIX), surprisingly little is known about the mechanism of its toxicity, aside from its phototoxicity. The cellular protein interactions of PPIX might provide insight into modulators of PPIX-induced cell death. Here we report the development of PPB, a biotin-conjugated, PPIX-probe that captures proteins capable of interacting with PPIX. Quantitative proteomics in a diverse panel of mammalian cell lines reveal a high degree of concordance for PPB-interacting proteins identified for each cell line. Most differences are quantitative, despite marked differences in PPIX formation and sensitivity. Pathway and quantitative difference analysis indicate that iron and heme metabolism proteins are prominent among PPB-bound proteins in fibroblasts, which undergo PPIX-mediated death determined to occur through ferroptosis. PPB proteomic data (available at PRIDE ProteomeXchange # PXD042631) reveal that redox proteins from PRDX family of glutathione peroxidases interact with PPIX. Targeted gene knockdown of the mitochondrial PRDX3, but not PRDX1 or 2, enhance PPIX-induced death in fibroblasts, an effect blocked by the radical-trapping antioxidant, ferrostatin-1. Increased PPIX formation and death was also observed in a T-lymphoblastoid ferrochelatase-deficient leukemia cell line, suggesting that PPIX elevation might serve as a potential strategy for killing certain leukemias. Nature Publishing Group UK 2023-06-24 /pmc/articles/PMC10290680/ /pubmed/37355765 http://dx.doi.org/10.1038/s42003-023-05024-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Lynch, John
Wang, Yao
Li, Yuxin
Kavdia, Kanisha
Fukuda, Yu
Ranjit, Sabina
Robinson, Camenzind G.
Grace, Christy R.
Xia, Youlin
Peng, Junmin
Schuetz, John D.
A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin
title A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin
title_full A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin
title_fullStr A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin
title_full_unstemmed A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin
title_short A PPIX-binding probe facilitates discovery of PPIX-induced cell death modulation by peroxiredoxin
title_sort ppix-binding probe facilitates discovery of ppix-induced cell death modulation by peroxiredoxin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10290680/
https://www.ncbi.nlm.nih.gov/pubmed/37355765
http://dx.doi.org/10.1038/s42003-023-05024-5
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