Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>

ABSTRACT l-Cysteine is essential for virtually all living organisms, from bacteria to higher eukaryotes. Besides having a role in the synthesis of virtually all proteins and of taurine, cysteamine, glutathione, and other redox-regulating proteins, l-cysteine has important functions under anaerobic/m...

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Autores principales:	Ghulam Jeelani, Dan Sato, Tomoyoshi Soga, Haruo Watanabe, Tomoyoshi Nozaki
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
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spelling	oai:doaj.org-article:86568b76de414a8fa73a70fdc1c477402021-11-15T15:47:03ZMass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>10.1128/mBio.01995-142150-7511https://doaj.org/article/86568b76de414a8fa73a70fdc1c477402014-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01995-14https://doaj.org/toc/2150-7511ABSTRACT l-Cysteine is essential for virtually all living organisms, from bacteria to higher eukaryotes. Besides having a role in the synthesis of virtually all proteins and of taurine, cysteamine, glutathione, and other redox-regulating proteins, l-cysteine has important functions under anaerobic/microaerophilic conditions. In anaerobic or microaerophilic protozoan parasites, such as Entamoeba histolytica, l-cysteine has been implicated in growth, attachment, survival, and protection from oxidative stress. However, a specific role of this amino acid or related metabolic intermediates is not well understood. In this study, using stable-isotope-labeled l-cysteine and capillary electrophoresis-time of flight mass spectrometry, we investigated the metabolism of l-cysteine in E. histolytica. [U-13C3, 15N]l-cysteine was rapidly metabolized into three unknown metabolites, besides l-cystine and l-alanine. These metabolites were identified as thiazolidine-4-carboxylic acid (T4C), 2-methyl thiazolidine-4-carboxylic acid (MT4C), and 2-ethyl-thiazolidine-4-carboxylic acid (ET4C), the condensation products of l-cysteine with aldehydes. We demonstrated that these 2-(R)-thiazolidine-4-carboxylic acids serve for storage of l-cysteine. Liberation of l-cysteine occurred when T4C was incubated with amebic lysates, suggesting enzymatic degradation of these l-cysteine derivatives. Furthermore, T4C and MT4C significantly enhanced trophozoite growth and reduced intracellular reactive oxygen species (ROS) levels when it was added to cultures, suggesting that 2-(R)-thiazolidine-4-carboxylic acids are involved in the defense against oxidative stress. IMPORTANCE Amebiasis is a human parasitic disease caused by the protozoan parasite Entamoeba histolytica. In this parasite, l-cysteine is the principal low-molecular-weight thiol and is assumed to play a significant role in supplying the amino acid during trophozoite invasion, particularly when the parasites move from the anaerobic intestinal lumen to highly oxygenated tissues in the intestine and the liver. It is well known that E. histolytica needs a comparatively high concentration of l-cysteine for its axenic cultivation. However, the reason for and the metabolic fate of l-cysteine in this parasite are not well understood. Here, using a metabolomic and stable-isotope-labeled approach, we investigated the metabolic fate of this amino acid in these parasites. We found that l-cysteine inside the cell rapidly reacts with aldehydes to form 2-(R)-thiazolidine-4-carboxylic acid. We showed that these 2-(R)-thiazolidine-4-carboxylic derivatives serve as an l-cysteine source, promote growth, and protect cells against oxidative stress by scavenging aldehydes and reducing the ROS level. Our findings represent the first demonstration of 2-(R)-thiazolidine-4-carboxylic acids and their roles in protozoan parasites.Ghulam JeelaniDan SatoTomoyoshi SogaHaruo WatanabeTomoyoshi NozakiAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 6 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Ghulam Jeelani Dan Sato Tomoyoshi Soga Haruo Watanabe Tomoyoshi Nozaki Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>
description	ABSTRACT l-Cysteine is essential for virtually all living organisms, from bacteria to higher eukaryotes. Besides having a role in the synthesis of virtually all proteins and of taurine, cysteamine, glutathione, and other redox-regulating proteins, l-cysteine has important functions under anaerobic/microaerophilic conditions. In anaerobic or microaerophilic protozoan parasites, such as Entamoeba histolytica, l-cysteine has been implicated in growth, attachment, survival, and protection from oxidative stress. However, a specific role of this amino acid or related metabolic intermediates is not well understood. In this study, using stable-isotope-labeled l-cysteine and capillary electrophoresis-time of flight mass spectrometry, we investigated the metabolism of l-cysteine in E. histolytica. [U-13C3, 15N]l-cysteine was rapidly metabolized into three unknown metabolites, besides l-cystine and l-alanine. These metabolites were identified as thiazolidine-4-carboxylic acid (T4C), 2-methyl thiazolidine-4-carboxylic acid (MT4C), and 2-ethyl-thiazolidine-4-carboxylic acid (ET4C), the condensation products of l-cysteine with aldehydes. We demonstrated that these 2-(R)-thiazolidine-4-carboxylic acids serve for storage of l-cysteine. Liberation of l-cysteine occurred when T4C was incubated with amebic lysates, suggesting enzymatic degradation of these l-cysteine derivatives. Furthermore, T4C and MT4C significantly enhanced trophozoite growth and reduced intracellular reactive oxygen species (ROS) levels when it was added to cultures, suggesting that 2-(R)-thiazolidine-4-carboxylic acids are involved in the defense against oxidative stress. IMPORTANCE Amebiasis is a human parasitic disease caused by the protozoan parasite Entamoeba histolytica. In this parasite, l-cysteine is the principal low-molecular-weight thiol and is assumed to play a significant role in supplying the amino acid during trophozoite invasion, particularly when the parasites move from the anaerobic intestinal lumen to highly oxygenated tissues in the intestine and the liver. It is well known that E. histolytica needs a comparatively high concentration of l-cysteine for its axenic cultivation. However, the reason for and the metabolic fate of l-cysteine in this parasite are not well understood. Here, using a metabolomic and stable-isotope-labeled approach, we investigated the metabolic fate of this amino acid in these parasites. We found that l-cysteine inside the cell rapidly reacts with aldehydes to form 2-(R)-thiazolidine-4-carboxylic acid. We showed that these 2-(R)-thiazolidine-4-carboxylic derivatives serve as an l-cysteine source, promote growth, and protect cells against oxidative stress by scavenging aldehydes and reducing the ROS level. Our findings represent the first demonstration of 2-(R)-thiazolidine-4-carboxylic acids and their roles in protozoan parasites.
format	article
author	Ghulam Jeelani Dan Sato Tomoyoshi Soga Haruo Watanabe Tomoyoshi Nozaki
author_facet	Ghulam Jeelani Dan Sato Tomoyoshi Soga Haruo Watanabe Tomoyoshi Nozaki
author_sort	Ghulam Jeelani
title	Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>
title_short	Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>
title_full	Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>
title_fullStr	Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>
title_full_unstemmed	Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>
title_sort	mass spectrometric analysis of <sc>l</sc>-cysteine metabolism: physiological role and fate of <sc>l</sc>-cysteine in the enteric protozoan parasite <named-content content-type="genus-species">entamoeba histolytica</named-content>
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/86568b76de414a8fa73a70fdc1c47740
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Mass Spectrometric Analysis of <sc>l</sc>-Cysteine Metabolism: Physiological Role and Fate of <sc>l</sc>-Cysteine in the Enteric Protozoan Parasite <named-content content-type="genus-species">Entamoeba histolytica</named-content>

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