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A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence

Here, we found a natural compound, embonic acid (EA), that can specifically inhibit the enzymatic activity of mitochondrial NAD(P)(+)-dependent malic enzyme (m-NAD(P)-ME, ME2) either in vitro or in vivo. The in vitro IC(50) value of EA for m-NAD(P)-ME was 1.4 ± 0.4 μM. Mutagenesis and binding studie...

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Autores principales: Hsieh, Ju-Yi, Li, Shao-Yu, Tsai, Wen-Chen, Liu, Jyung-Hurng, Lin, Chih-Li, Liu, Guang-Yaw, Hung, Hui-Chih
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652989/
https://www.ncbi.nlm.nih.gov/pubmed/26008970
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author Hsieh, Ju-Yi
Li, Shao-Yu
Tsai, Wen-Chen
Liu, Jyung-Hurng
Lin, Chih-Li
Liu, Guang-Yaw
Hung, Hui-Chih
author_facet Hsieh, Ju-Yi
Li, Shao-Yu
Tsai, Wen-Chen
Liu, Jyung-Hurng
Lin, Chih-Li
Liu, Guang-Yaw
Hung, Hui-Chih
author_sort Hsieh, Ju-Yi
collection PubMed
description Here, we found a natural compound, embonic acid (EA), that can specifically inhibit the enzymatic activity of mitochondrial NAD(P)(+)-dependent malic enzyme (m-NAD(P)-ME, ME2) either in vitro or in vivo. The in vitro IC(50) value of EA for m-NAD(P)-ME was 1.4 ± 0.4 μM. Mutagenesis and binding studies revealed that the putative binding site of EA on m-NAD(P)-ME is located at the fumarate binding site or at the dimer interface near the site. Inhibition studies reveal that EA displayed a non-competitive inhibition pattern, which demonstrated that the binding site of EA was distinct from the active site of the enzyme. Therefore, EA is thought to be an allosteric inhibitor of m-NAD(P)-ME. Both EA treatment and knockdown of m-NAD(P)-ME by shRNA inhibited the growth of H1299 cancer cells. The protein expression and mRNA synthesis of m-NAD(P)-ME in H1299 cells were not influenced by EA, suggesting that the EA-inhibited H1299 cell growth occurs through the suppression of in vivo m-NAD(P)-ME activity EA treatment further induced the cellular senescence of H1299 cells. However, down-regulation of the enzyme-induced cellular senescence was not through p53. Therefore, the EA-evoked senescence of H1299 cells may occur directly through the inhibition of ME2 or a p53-independent pathway.
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spelling pubmed-46529892015-12-02 A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence Hsieh, Ju-Yi Li, Shao-Yu Tsai, Wen-Chen Liu, Jyung-Hurng Lin, Chih-Li Liu, Guang-Yaw Hung, Hui-Chih Oncotarget Research Paper Here, we found a natural compound, embonic acid (EA), that can specifically inhibit the enzymatic activity of mitochondrial NAD(P)(+)-dependent malic enzyme (m-NAD(P)-ME, ME2) either in vitro or in vivo. The in vitro IC(50) value of EA for m-NAD(P)-ME was 1.4 ± 0.4 μM. Mutagenesis and binding studies revealed that the putative binding site of EA on m-NAD(P)-ME is located at the fumarate binding site or at the dimer interface near the site. Inhibition studies reveal that EA displayed a non-competitive inhibition pattern, which demonstrated that the binding site of EA was distinct from the active site of the enzyme. Therefore, EA is thought to be an allosteric inhibitor of m-NAD(P)-ME. Both EA treatment and knockdown of m-NAD(P)-ME by shRNA inhibited the growth of H1299 cancer cells. The protein expression and mRNA synthesis of m-NAD(P)-ME in H1299 cells were not influenced by EA, suggesting that the EA-inhibited H1299 cell growth occurs through the suppression of in vivo m-NAD(P)-ME activity EA treatment further induced the cellular senescence of H1299 cells. However, down-regulation of the enzyme-induced cellular senescence was not through p53. Therefore, the EA-evoked senescence of H1299 cells may occur directly through the inhibition of ME2 or a p53-independent pathway. Impact Journals LLC 2015-05-19 /pmc/articles/PMC4652989/ /pubmed/26008970 Text en Copyright: © 2015 Hsieh et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Hsieh, Ju-Yi
Li, Shao-Yu
Tsai, Wen-Chen
Liu, Jyung-Hurng
Lin, Chih-Li
Liu, Guang-Yaw
Hung, Hui-Chih
A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence
title A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence
title_full A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence
title_fullStr A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence
title_full_unstemmed A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence
title_short A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)(+)-dependent malic enzyme (ME2) and induces cellular senescence
title_sort small-molecule inhibitor suppresses the tumor-associated mitochondrial nad(p)(+)-dependent malic enzyme (me2) and induces cellular senescence
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652989/
https://www.ncbi.nlm.nih.gov/pubmed/26008970
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