{"id":31602,"date":"1997-01-01T00:00:00","date_gmt":"1997-01-01T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/expresion-de-la-s-adenosil-l-metionina-sintetasa-de-higado-de-rata-en-escherichia-coli-y-analisis-del-papel-estructural-y-funcional-de-sus-grupos-sulfhidrilo-mediante-mutagenesis-dirigida\/"},"modified":"1997-01-01T00:00:00","modified_gmt":"1997-01-01T00:00:00","slug":"expresion-de-la-s-adenosil-l-metionina-sintetasa-de-higado-de-rata-en-escherichia-coli-y-analisis-del-papel-estructural-y-funcional-de-sus-grupos-sulfhidrilo-mediante-mutagenesis-dirigida","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/quimica\/expresion-de-la-s-adenosil-l-metionina-sintetasa-de-higado-de-rata-en-escherichia-coli-y-analisis-del-papel-estructural-y-funcional-de-sus-grupos-sulfhidrilo-mediante-mutagenesis-dirigida\/","title":{"rendered":"Expresion de la s-adenosil-l-metionina sintetasa de higado de rata en escherichia coli y analisis del papel estructural y funcional de sus grupos sulfhidrilo mediante mutagenesis dirigida."},"content":{"rendered":"<h2>Tesis doctoral de <strong> Jes\u00fas Mingorance Cruz <\/strong><\/h2>\n<p>La region codificante de la s-adenosil-l-metionina sintetasa de higado de rata fue subcionada en un vector de expresion en e.Coli. Este sistema produjo gran cantidad de enzima recombinante, la mayor parte en forma de cuerpos de inclusion, aunque tambien se obtuvo una fraccion en forma soluble. La enzima soluble era activa, y formaba tetrameros y dimeros. Se desarrollo un protocolo para purificarla, y se caracterizo la enzima obtenida. Esta contenia una mezcla en equilibrio de dos isoformas, tetrameros y dimeros, con una constante de equilibrio aparente de aproximadamente 5&#215;10 5 m-1; el tetramero se disociaba siguiendo una cinetica de primer orden, con una velocidad de k-1= (8.1 +-0.4) x10 -4 s -1 y una vida media de unos 14 minutos a 4 grados c. La actividad de la enzima recombinante era inhibida de forma dependiente de dosis por no, h2o2 y gssg, con constantes de inhibicion de 0.08, 0.25 y 2 mm respectivamente. Para estudiar el papel estructural y funcional de los grupos sulfhidrilo de la enzima se construyeron diez mutantes, en cada uno de los cuales se cambio uno de los codones de cisteina por uno de serina. Todos los mutantes produjeron proteina soluble cuando se expresaron en e. Coli. Todos tenian actividad s-adenosil-l-metionina sintetasa, y todos producian dos isoformas, sin embargo, la cantidad relativa de cada una vario en algunos casos: las mutaciones c35s, c57s, c61s y c105s produjeron un aumento de la cantidad relativa de tetramero, mientras que por el contrario la mutacion c69s produjo un aumento de la cantidad de dimero. La sensibilidad de los mutantes a los agentes oxidantes no, h2o2 y gssg vario en cada caso, siendo el mas notable el mutante c121s, que era insensible al no y al h2o2, y poco sensible al gssg.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Expresion de la s-adenosil-l-metionina sintetasa de higado de rata en escherichia coli y analisis del papel estructural y funcional de sus grupos sulfhidrilo mediante mutagenesis dirigida.<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Expresion de la s-adenosil-l-metionina sintetasa de higado de rata en escherichia coli y analisis del papel estructural y funcional de sus grupos sulfhidrilo mediante mutagenesis dirigida. <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Jes\u00fas Mingorance Cruz <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Aut\u00f3noma de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 01\/01\/1997<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li> Pajares Tarancon M. Angeles<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: Ernest Giralt Lledo <\/li>\n<li>Lisardo Bosc\u00e1 Gomar (vocal)<\/li>\n<li>Jos\u00e9 Mar\u00eda Mato De La Paz (vocal)<\/li>\n<li>\u00c1lvaro Martinez  Del Pozo (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Jes\u00fas Mingorance Cruz La region codificante de la s-adenosil-l-metionina sintetasa de higado de rata fue subcionada en [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"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-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-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-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-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-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-gradient":""}},"footnotes":""},"categories":[160,115,17],"tags":[87918,1100,85696,14571,5664,14570],"class_list":["post-31602","post","type-post","status-publish","format-standard","hentry","category-bioquimica","category-bioquimica-molecular","category-quimica","tag-alvaro-Martinez-del-pozo","tag-ernest-giralt-lledo","tag-jesus-mingorance-cruz","tag-jose-maria-mato-de-la-paz","tag-lisardo-bosca-gomar","tag-pajares-tarancon-m-angeles"],"_links":{"self":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts\/31602","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/comments?post=31602"}],"version-history":[{"count":0,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts\/31602\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/media?parent=31602"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/categories?post=31602"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/tags?post=31602"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}