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Development of Chemical Proteomics for the Folateome and Analysis of the Kinetoplastid Folateome
[Image: see text] The folate pathway has been extensively studied in a number of organisms, with its essentiality exploited by a number of drugs. However, there has been little success in developing drugs that target folate metabolism in the kinetoplastids. Despite compounds being identified which s...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6199744/ https://www.ncbi.nlm.nih.gov/pubmed/30264983 http://dx.doi.org/10.1021/acsinfecdis.8b00097 |
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author | Webster, Lauren A. Thomas, Michael Urbaniak, Michael Wyllie, Susan Ong, Han Tinti, Michele Fairlamb, Alan H. Boesche, Markus Ghidelli-Disse, Sonja Drewes, Gerard Gilbert, Ian H. |
author_facet | Webster, Lauren A. Thomas, Michael Urbaniak, Michael Wyllie, Susan Ong, Han Tinti, Michele Fairlamb, Alan H. Boesche, Markus Ghidelli-Disse, Sonja Drewes, Gerard Gilbert, Ian H. |
author_sort | Webster, Lauren A. |
collection | PubMed |
description | [Image: see text] The folate pathway has been extensively studied in a number of organisms, with its essentiality exploited by a number of drugs. However, there has been little success in developing drugs that target folate metabolism in the kinetoplastids. Despite compounds being identified which show significant inhibition of the parasite enzymes, this activity does not translate well into cellular and animal models of disease. Understanding to which enzymes antifolates bind under physiological conditions and how this corresponds to the phenotypic response could provide insight on how to target the folate pathway in these organisms. To facilitate this, we have adopted a chemical proteomics approach to study binding of compounds to enzymes of folate metabolism. Clinical and literature antifolate compounds were immobilized onto resins to allow for “pull down” of the proteins in the “folateome”. Using competition studies, proteins, which bind the beads specifically and nonspecifically, were identified in parasite lysate (Trypanosoma brucei and Leishmania major) for each antifolate compound. Proteins were identified through tryptic digest, tandem mass tag (TMT) labeling of peptides followed by LC-MS/MS. This approach was further exploited by creating a combined folate resin (folate beads). The resin could pull down up to 9 proteins from the folateome. This information could be exploited in gaining a better understanding of folate metabolism in kinetoplastids and other organisms. |
format | Online Article Text |
id | pubmed-6199744 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-61997442018-11-06 Development of Chemical Proteomics for the Folateome and Analysis of the Kinetoplastid Folateome Webster, Lauren A. Thomas, Michael Urbaniak, Michael Wyllie, Susan Ong, Han Tinti, Michele Fairlamb, Alan H. Boesche, Markus Ghidelli-Disse, Sonja Drewes, Gerard Gilbert, Ian H. ACS Infect Dis [Image: see text] The folate pathway has been extensively studied in a number of organisms, with its essentiality exploited by a number of drugs. However, there has been little success in developing drugs that target folate metabolism in the kinetoplastids. Despite compounds being identified which show significant inhibition of the parasite enzymes, this activity does not translate well into cellular and animal models of disease. Understanding to which enzymes antifolates bind under physiological conditions and how this corresponds to the phenotypic response could provide insight on how to target the folate pathway in these organisms. To facilitate this, we have adopted a chemical proteomics approach to study binding of compounds to enzymes of folate metabolism. Clinical and literature antifolate compounds were immobilized onto resins to allow for “pull down” of the proteins in the “folateome”. Using competition studies, proteins, which bind the beads specifically and nonspecifically, were identified in parasite lysate (Trypanosoma brucei and Leishmania major) for each antifolate compound. Proteins were identified through tryptic digest, tandem mass tag (TMT) labeling of peptides followed by LC-MS/MS. This approach was further exploited by creating a combined folate resin (folate beads). The resin could pull down up to 9 proteins from the folateome. This information could be exploited in gaining a better understanding of folate metabolism in kinetoplastids and other organisms. American Chemical Society 2018-09-28 2018-10-12 /pmc/articles/PMC6199744/ /pubmed/30264983 http://dx.doi.org/10.1021/acsinfecdis.8b00097 Text en Copyright © 2018 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. |
spellingShingle | Webster, Lauren A. Thomas, Michael Urbaniak, Michael Wyllie, Susan Ong, Han Tinti, Michele Fairlamb, Alan H. Boesche, Markus Ghidelli-Disse, Sonja Drewes, Gerard Gilbert, Ian H. Development of Chemical Proteomics for the Folateome and Analysis of the Kinetoplastid Folateome |
title | Development of Chemical Proteomics for the Folateome
and Analysis of the Kinetoplastid Folateome |
title_full | Development of Chemical Proteomics for the Folateome
and Analysis of the Kinetoplastid Folateome |
title_fullStr | Development of Chemical Proteomics for the Folateome
and Analysis of the Kinetoplastid Folateome |
title_full_unstemmed | Development of Chemical Proteomics for the Folateome
and Analysis of the Kinetoplastid Folateome |
title_short | Development of Chemical Proteomics for the Folateome
and Analysis of the Kinetoplastid Folateome |
title_sort | development of chemical proteomics for the folateome
and analysis of the kinetoplastid folateome |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6199744/ https://www.ncbi.nlm.nih.gov/pubmed/30264983 http://dx.doi.org/10.1021/acsinfecdis.8b00097 |
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