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Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus

Lecithin-dependent thermolabile hemolysin (LDH) is a virulence factor excreted by Vibrio parahaemolyticus, a marine bacterium that causes important losses in shrimp farming. In this study, the function of LDH was investigated through its inhibition by metal ions (Mg(2+), Ca(2+), Mn(2+), Co(2+), Ni(2...

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Autores principales: Vazquez-Armenta, Francisco Javier, Valdez-Olmos, Uriel Felipe, Arvizu-Flores, Aldo Alejandro, Ayala-Zavala, Jesus Fernando, Ochoa-Leyva, Adrian, Lopez-Zavala, Alonso Alexis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9506434/
https://www.ncbi.nlm.nih.gov/pubmed/36136547
http://dx.doi.org/10.3390/toxins14090609
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author Vazquez-Armenta, Francisco Javier
Valdez-Olmos, Uriel Felipe
Arvizu-Flores, Aldo Alejandro
Ayala-Zavala, Jesus Fernando
Ochoa-Leyva, Adrian
Lopez-Zavala, Alonso Alexis
author_facet Vazquez-Armenta, Francisco Javier
Valdez-Olmos, Uriel Felipe
Arvizu-Flores, Aldo Alejandro
Ayala-Zavala, Jesus Fernando
Ochoa-Leyva, Adrian
Lopez-Zavala, Alonso Alexis
author_sort Vazquez-Armenta, Francisco Javier
collection PubMed
description Lecithin-dependent thermolabile hemolysin (LDH) is a virulence factor excreted by Vibrio parahaemolyticus, a marine bacterium that causes important losses in shrimp farming. In this study, the function of LDH was investigated through its inhibition by metal ions (Mg(2+), Ca(2+), Mn(2+), Co(2+), Ni(2+) and Cu(2+)) and chemical modification reagents: β-mercaptoethanol (βME), phenylmethylsulfonyl fluoride (PMSF) and diethyl pyrocarbonate (DEPC). LDH was expressed in the Escherichia coli strain BL-21, purified under denaturing conditions, and the enzymatic activity was evaluated. Cu(2+), Ni(2+), Co(2+) and Ca(2+) at 1 mmol/L inhibited the LDH esterase activity by 20–95%, while Mg(2+) and Mn(2+) slightly increased its activity. Additionally, PMSF and DEPC at 1 mmol/L inhibited the enzymatic activity by 40% and 80%, respectively. Dose-response analysis showed that DEPC was the best-evaluated inhibitor (IC(50) = 0.082 mmol/L), followed by Cu(2+) > Co(2+) > Ni(2+) and PMSF (IC(50) = 0.146–1.5 mmol/L). Multiple sequence alignment of LDH of V. parahaemolyticus against other Vibrio species showed that LDH has well-conserved GDSL and SGNH motifs, characteristic of the hydrolase/esterase superfamily. Additionally, the homology model showed that the conserved catalytic triad His-Ser-Asp was in the LDH active site. Our results showed that the enzymatic activity of LDH from V. parahaemolyticus was modulated by metal ions and chemical modification, which could be related to the interaction with catalytic amino acid residues such as Ser153 and/or His 393.
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spelling pubmed-95064342022-09-24 Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus Vazquez-Armenta, Francisco Javier Valdez-Olmos, Uriel Felipe Arvizu-Flores, Aldo Alejandro Ayala-Zavala, Jesus Fernando Ochoa-Leyva, Adrian Lopez-Zavala, Alonso Alexis Toxins (Basel) Article Lecithin-dependent thermolabile hemolysin (LDH) is a virulence factor excreted by Vibrio parahaemolyticus, a marine bacterium that causes important losses in shrimp farming. In this study, the function of LDH was investigated through its inhibition by metal ions (Mg(2+), Ca(2+), Mn(2+), Co(2+), Ni(2+) and Cu(2+)) and chemical modification reagents: β-mercaptoethanol (βME), phenylmethylsulfonyl fluoride (PMSF) and diethyl pyrocarbonate (DEPC). LDH was expressed in the Escherichia coli strain BL-21, purified under denaturing conditions, and the enzymatic activity was evaluated. Cu(2+), Ni(2+), Co(2+) and Ca(2+) at 1 mmol/L inhibited the LDH esterase activity by 20–95%, while Mg(2+) and Mn(2+) slightly increased its activity. Additionally, PMSF and DEPC at 1 mmol/L inhibited the enzymatic activity by 40% and 80%, respectively. Dose-response analysis showed that DEPC was the best-evaluated inhibitor (IC(50) = 0.082 mmol/L), followed by Cu(2+) > Co(2+) > Ni(2+) and PMSF (IC(50) = 0.146–1.5 mmol/L). Multiple sequence alignment of LDH of V. parahaemolyticus against other Vibrio species showed that LDH has well-conserved GDSL and SGNH motifs, characteristic of the hydrolase/esterase superfamily. Additionally, the homology model showed that the conserved catalytic triad His-Ser-Asp was in the LDH active site. Our results showed that the enzymatic activity of LDH from V. parahaemolyticus was modulated by metal ions and chemical modification, which could be related to the interaction with catalytic amino acid residues such as Ser153 and/or His 393. MDPI 2022-09-01 /pmc/articles/PMC9506434/ /pubmed/36136547 http://dx.doi.org/10.3390/toxins14090609 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Vazquez-Armenta, Francisco Javier
Valdez-Olmos, Uriel Felipe
Arvizu-Flores, Aldo Alejandro
Ayala-Zavala, Jesus Fernando
Ochoa-Leyva, Adrian
Lopez-Zavala, Alonso Alexis
Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus
title Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus
title_full Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus
title_fullStr Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus
title_full_unstemmed Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus
title_short Metal Ions and Chemical Modification Reagents Inhibit the Enzymatic Activity of Lecithin-Dependent Hemolysin from Vibrio parahaemolyticus
title_sort metal ions and chemical modification reagents inhibit the enzymatic activity of lecithin-dependent hemolysin from vibrio parahaemolyticus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9506434/
https://www.ncbi.nlm.nih.gov/pubmed/36136547
http://dx.doi.org/10.3390/toxins14090609
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