{"id":659,"date":"2023-12-11T11:43:00","date_gmt":"2023-12-11T14:43:00","guid":{"rendered":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659"},"modified":"2024-04-04T11:46:35","modified_gmt":"2024-04-04T14:46:35","slug":"from-in-silico-screening-to-in-vitro-evaluation-enhancing-the-detection-ofmicrocystins-with-engineered-pp1-mutant-variants","status":"publish","type":"post","link":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659","title":{"rendered":"From in-silico screening to in-vitro evaluation: Enhancing the detection ofMicrocystins with engineered PP1 mutant variants"},"content":{"rendered":"\n<p>Ezequiel J. Alba-Posse, Carlos David Bruque, Yamila Gandola, Javier Gasulla, Alejandro D. Nadra<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Abstract<\/h2>\n\n\n\n<p>Cyanotoxins produced during harmful algal blooms (CyanoHABs) have become a worldwide issue of concern. Microcystins (MC) are the most ubiquitous group of cyanotoxins and have known carcinogenic and hepatotoxic effects. The protein phosphatase inhibition assays (PPIAs), based on the inhibition of Protein Phosphatase 1\/2A (PP1\/PP2A) by MC, are one of the most cost-effective options for detecting MC. In this work, we aimed to design in-silico and evaluate in-vitro mutant variants of the PP1 protein, in order to enhance their capabilities as a MC biosensor. To this end, we performed an in-silico active site-saturated mutagenesis screening, followed by stability and docking affinity calculation with the MCLR cyanotoxin. Candidates with improved both affinity and stability were further tested in a fully flexible active-site docking. The best-scored mutations (19) were individually analysed regarding their locations and interactions. Four of them (p.D197F; p.Q249Y; p.S129W; p.D220Q) were selected for in-vitro expression and evaluation. Mutant p.D197F, exhibited a significant increment in inhibition by MCLR with respect to the WT, while showing a non-significant difference in stability nor activity. This successful PP1 inhibition enhancement suggests the potential of the p.D197F variant for practical MC detection applications.<\/p>\n\n\n\n<div class=\"wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button has-custom-width wp-block-button__width-100\"><a class=\"wp-block-button__link wp-element-button\" href=\"https:\/\/www.sciencedirect.com\/science\/article\/abs\/pii\/S1047847723001065?via%3Dihub\" target=\"_blank\" rel=\"noreferrer noopener\">Ver Publicaci\u00f3n<\/a><\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Ezequiel J. Alba-Posse, Carlos David Bruque, Yamila Gandola, Javier Gasulla, Alejandro D. Nadra Abstract Cyanotoxins produced during harmful algal blooms (CyanoHABs) have become a worldwide issue of concern. Microcystins (MC) are the most ubiquitous group of cyanotoxins and have known carcinogenic and hepatotoxic effects. The protein phosphatase inhibition assays (PPIAs), based on the inhibition of [&hellip;]<\/p>\n","protected":false},"author":2,"featured_media":521,"comment_status":"closed","ping_status":"","sticky":false,"template":"","format":"standard","meta":{"_eb_attr":"","_uag_custom_page_level_css":"","advgb_blocks_editor_width":"","advgb_blocks_columns_visual_guide":"","site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"categories":[11],"tags":[],"class_list":["post-659","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-publicaciones"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v23.3 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>From in-silico screening to in-vitro evaluation: Enhancing the detection ofMicrocystins with engineered PP1 mutant variants - UCT HP<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659\" \/>\n<meta property=\"og:locale\" content=\"es_ES\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"From in-silico screening to in-vitro evaluation: Enhancing the detection ofMicrocystins with engineered PP1 mutant variants - UCT HP\" \/>\n<meta property=\"og:description\" content=\"Ezequiel J. Alba-Posse, Carlos David Bruque, Yamila Gandola, Javier Gasulla, Alejandro D. Nadra Abstract Cyanotoxins produced during harmful algal blooms (CyanoHABs) have become a worldwide issue of concern. Microcystins (MC) are the most ubiquitous group of cyanotoxins and have known carcinogenic and hepatotoxic effects. The protein phosphatase inhibition assays (PPIAs), based on the inhibition of [&hellip;]\" \/>\n<meta property=\"og:url\" content=\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659\" \/>\n<meta property=\"og:site_name\" content=\"UCT HP\" \/>\n<meta property=\"article:published_time\" content=\"2023-12-11T14:43:00+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2024-04-04T14:46:35+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/wp-content\/uploads\/2023\/10\/elsevier.png\" \/>\n\t<meta property=\"og:image:width\" content=\"250\" \/>\n\t<meta property=\"og:image:height\" content=\"200\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/png\" \/>\n<meta name=\"author\" content=\"Mirtha\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Escrito por\" \/>\n\t<meta name=\"twitter:data1\" content=\"Mirtha\" \/>\n\t<meta name=\"twitter:label2\" content=\"Tiempo de lectura\" \/>\n\t<meta name=\"twitter:data2\" content=\"1 minuto\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659#article\",\"isPartOf\":{\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659\"},\"author\":{\"name\":\"Mirtha\",\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/#\/schema\/person\/cb3d81609139435146e1257936dc9b98\"},\"headline\":\"From in-silico screening to in-vitro evaluation: Enhancing the detection ofMicrocystins with engineered PP1 mutant variants\",\"datePublished\":\"2023-12-11T14:43:00+00:00\",\"dateModified\":\"2024-04-04T14:46:35+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659\"},\"wordCount\":234,\"publisher\":{\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/#organization\"},\"image\":{\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659#primaryimage\"},\"thumbnailUrl\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/wp-content\/uploads\/2023\/10\/elsevier.png\",\"articleSection\":[\"publicaciones\"],\"inLanguage\":\"es\"},{\"@type\":\"WebPage\",\"@id\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659\",\"url\":\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?p=659\",\"name\":\"From in-silico screening to in-vitro evaluation: Enhancing the detection ofMicrocystins with engineered PP1 mutant variants - 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Alba-Posse, Carlos David Bruque, Yamila Gandola, Javier Gasulla, Alejandro D. Nadra Abstract Cyanotoxins produced during harmful algal blooms (CyanoHABs) have become a worldwide issue of concern. Microcystins (MC) are the most ubiquitous group of cyanotoxins and have known carcinogenic and hepatotoxic effects. The protein phosphatase inhibition assays (PPIAs), based on the inhibition of&hellip;","coauthors":[],"tax_additional":{"categories":{"linked":["<a href=\"https:\/\/ucthp.hospitalelcalafate.gob.ar\/?cat=11\" class=\"advgb-post-tax-term\">publicaciones<\/a>"],"unlinked":["<span class=\"advgb-post-tax-term\">publicaciones<\/span>"]}},"comment_count":"0","relative_dates":{"created":"Publicado 2 a\u00f1os hace","modified":"Actualizado 2 a\u00f1os hace"},"absolute_dates":{"created":"Publicado el 11 de diciembre de 2023","modified":"Actualizado el 4 de abril de 2024"},"absolute_dates_time":{"created":"Publicado el 11 de diciembre de 2023 11:43 am","modified":"Actualizado el 4 de abril de 2024 11:46 am"},"featured_img_caption":"","series_order":"","_links":{"self":[{"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/posts\/659","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=659"}],"version-history":[{"count":2,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/posts\/659\/revisions"}],"predecessor-version":[{"id":661,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/posts\/659\/revisions\/661"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=\/wp\/v2\/media\/521"}],"wp:attachment":[{"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=659"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=659"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/ucthp.hospitalelcalafate.gob.ar\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=659"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}