Cargando…

Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome

Pyridoxal 5′-phosphate (PLP) is a versatile cofactor that catalyzes a plethora of chemical transformations within a cell. Although many human PLP-dependent enzymes (PLP-DEs) with crucial physiological and pathological roles are known, a global method enabling their cellular profiling is lacking. Her...

Descripción completa

Detalles Bibliográficos
Autores principales: Fux, Anja, Pfanzelt, Martin, Kirsch, Volker C., Hoegl, Annabelle, Sieber, Stephan A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6876276/
https://www.ncbi.nlm.nih.gov/pubmed/31447350
http://dx.doi.org/10.1016/j.chembiol.2019.08.003
_version_ 1783473182130307072
author Fux, Anja
Pfanzelt, Martin
Kirsch, Volker C.
Hoegl, Annabelle
Sieber, Stephan A.
author_facet Fux, Anja
Pfanzelt, Martin
Kirsch, Volker C.
Hoegl, Annabelle
Sieber, Stephan A.
author_sort Fux, Anja
collection PubMed
description Pyridoxal 5′-phosphate (PLP) is a versatile cofactor that catalyzes a plethora of chemical transformations within a cell. Although many human PLP-dependent enzymes (PLP-DEs) with crucial physiological and pathological roles are known, a global method enabling their cellular profiling is lacking. Here, we demonstrate the utility of a cofactor probe for the identification of human PLP-binding proteins in living cells. Striking selectivity of human pyridoxal kinase led to a customized labeling strategy covering a large fraction of known PLP-binding proteins across various cancer-derived cell lines. Labeling intensities of some PLP-DEs varied depending on the cell type while the overall protein expression levels of these proteins remained constant. In addition, we applied the methodology for in situ screening of PLP-antagonists and unraveled known binders as well as unknown off-targets. Taken together, our proteome-wide method to study PLP-DEs in human cancer-derived cells enables global understanding of the interactome of this important cofactor.
format Online
Article
Text
id pubmed-6876276
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Cell Press
record_format MEDLINE/PubMed
spelling pubmed-68762762019-11-29 Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome Fux, Anja Pfanzelt, Martin Kirsch, Volker C. Hoegl, Annabelle Sieber, Stephan A. Cell Chem Biol Article Pyridoxal 5′-phosphate (PLP) is a versatile cofactor that catalyzes a plethora of chemical transformations within a cell. Although many human PLP-dependent enzymes (PLP-DEs) with crucial physiological and pathological roles are known, a global method enabling their cellular profiling is lacking. Here, we demonstrate the utility of a cofactor probe for the identification of human PLP-binding proteins in living cells. Striking selectivity of human pyridoxal kinase led to a customized labeling strategy covering a large fraction of known PLP-binding proteins across various cancer-derived cell lines. Labeling intensities of some PLP-DEs varied depending on the cell type while the overall protein expression levels of these proteins remained constant. In addition, we applied the methodology for in situ screening of PLP-antagonists and unraveled known binders as well as unknown off-targets. Taken together, our proteome-wide method to study PLP-DEs in human cancer-derived cells enables global understanding of the interactome of this important cofactor. Cell Press 2019-10-17 /pmc/articles/PMC6876276/ /pubmed/31447350 http://dx.doi.org/10.1016/j.chembiol.2019.08.003 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Fux, Anja
Pfanzelt, Martin
Kirsch, Volker C.
Hoegl, Annabelle
Sieber, Stephan A.
Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome
title Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome
title_full Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome
title_fullStr Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome
title_full_unstemmed Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome
title_short Customizing Functionalized Cofactor Mimics to Study the Human Pyridoxal 5′-Phosphate-Binding Proteome
title_sort customizing functionalized cofactor mimics to study the human pyridoxal 5′-phosphate-binding proteome
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6876276/
https://www.ncbi.nlm.nih.gov/pubmed/31447350
http://dx.doi.org/10.1016/j.chembiol.2019.08.003
work_keys_str_mv AT fuxanja customizingfunctionalizedcofactormimicstostudythehumanpyridoxal5phosphatebindingproteome
AT pfanzeltmartin customizingfunctionalizedcofactormimicstostudythehumanpyridoxal5phosphatebindingproteome
AT kirschvolkerc customizingfunctionalizedcofactormimicstostudythehumanpyridoxal5phosphatebindingproteome
AT hoeglannabelle customizingfunctionalizedcofactormimicstostudythehumanpyridoxal5phosphatebindingproteome
AT sieberstephana customizingfunctionalizedcofactormimicstostudythehumanpyridoxal5phosphatebindingproteome