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2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo
Pyridoxal 5′-phosphate (PLP) is an essential cofactor for a class of enzymes that catalyze diverse reactions in central metabolism. The catalytic mechanism of some PLP-dependent enzymes involves the generation of reactive enamine intermediates like 2-aminoacrylate (2AA). 2AA can covalently modify PL...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9127364/ https://www.ncbi.nlm.nih.gov/pubmed/35460692 http://dx.doi.org/10.1016/j.jbc.2022.101970 |
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author | Shen, Wangchen Borchert, Andrew J. Downs, Diana M. |
author_facet | Shen, Wangchen Borchert, Andrew J. Downs, Diana M. |
author_sort | Shen, Wangchen |
collection | PubMed |
description | Pyridoxal 5′-phosphate (PLP) is an essential cofactor for a class of enzymes that catalyze diverse reactions in central metabolism. The catalytic mechanism of some PLP-dependent enzymes involves the generation of reactive enamine intermediates like 2-aminoacrylate (2AA). 2AA can covalently modify PLP in the active site of some PLP-dependent enzymes and subsequently inactivate the enzyme through the formation of a PLP–pyruvate adduct. In the absence of the enamine/imine deaminase RidA, Salmonella enterica experiences 2AA-mediated metabolic stress. Surprisingly, PLP-dependent enzymes that generate endogenous 2AA appear to be immune to its attack, while other PLP-dependent enzymes accumulate damage in the presence of 2AA stress; however, structural determinants of 2AA sensitivity are unclear. In this study, we refined a molecular method to query proteins from diverse systems for their sensitivity to 2AA in vivo. This method was then used to examine active site residues of Alr, a 2AA-sensitive PLP-dependent enzyme, that affect its sensitivity to 2AA in vivo. Unexpectedly, our data also showed that a low level of 2AA stress can persist even in the presence of a functional RidA. In summary, this study expands our understanding of 2AA metabolism and takes an initial step toward characterizing the structural determinants influencing enzyme susceptibility to damage by free 2AA. |
format | Online Article Text |
id | pubmed-9127364 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-91273642022-05-25 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo Shen, Wangchen Borchert, Andrew J. Downs, Diana M. J Biol Chem Research Article Pyridoxal 5′-phosphate (PLP) is an essential cofactor for a class of enzymes that catalyze diverse reactions in central metabolism. The catalytic mechanism of some PLP-dependent enzymes involves the generation of reactive enamine intermediates like 2-aminoacrylate (2AA). 2AA can covalently modify PLP in the active site of some PLP-dependent enzymes and subsequently inactivate the enzyme through the formation of a PLP–pyruvate adduct. In the absence of the enamine/imine deaminase RidA, Salmonella enterica experiences 2AA-mediated metabolic stress. Surprisingly, PLP-dependent enzymes that generate endogenous 2AA appear to be immune to its attack, while other PLP-dependent enzymes accumulate damage in the presence of 2AA stress; however, structural determinants of 2AA sensitivity are unclear. In this study, we refined a molecular method to query proteins from diverse systems for their sensitivity to 2AA in vivo. This method was then used to examine active site residues of Alr, a 2AA-sensitive PLP-dependent enzyme, that affect its sensitivity to 2AA in vivo. Unexpectedly, our data also showed that a low level of 2AA stress can persist even in the presence of a functional RidA. In summary, this study expands our understanding of 2AA metabolism and takes an initial step toward characterizing the structural determinants influencing enzyme susceptibility to damage by free 2AA. American Society for Biochemistry and Molecular Biology 2022-04-20 /pmc/articles/PMC9127364/ /pubmed/35460692 http://dx.doi.org/10.1016/j.jbc.2022.101970 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Shen, Wangchen Borchert, Andrew J. Downs, Diana M. 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo |
title | 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo |
title_full | 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo |
title_fullStr | 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo |
title_full_unstemmed | 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo |
title_short | 2-Aminoacrylate stress damages diverse PLP-dependent enzymes in vivo |
title_sort | 2-aminoacrylate stress damages diverse plp-dependent enzymes in vivo |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9127364/ https://www.ncbi.nlm.nih.gov/pubmed/35460692 http://dx.doi.org/10.1016/j.jbc.2022.101970 |
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