{"id":81450,"date":"2018-03-10T00:05:22","date_gmt":"2018-03-10T00:05:22","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/analisis-de-dominios-funcionales-del-transportador-abc-ycf1p-mediante-supresion-intragenica-y-mutagenesis-dirigida\/"},"modified":"2018-03-10T00:05:22","modified_gmt":"2018-03-10T00:05:22","slug":"analisis-de-dominios-funcionales-del-transportador-abc-ycf1p-mediante-supresion-intragenica-y-mutagenesis-dirigida","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/quimica\/analisis-de-dominios-funcionales-del-transportador-abc-ycf1p-mediante-supresion-intragenica-y-mutagenesis-dirigida\/","title":{"rendered":"An\u00e1lisis de dominios funcionales del transportador abc, ycf1p, mediante supresi\u00f3n intragenica y mutagenesis dirigida."},"content":{"rendered":"<h2>Tesis doctoral de <strong> Juan  Manuel Falcon Perez <\/strong><\/h2>\n<p>La prote\u00edna vacuolar ycf1p (yeast cadmium factor) est\u00e1 implicada en la resistencia a cd2+ y a otras sustancias t\u00f3xicas que son conjutadas con glutation en saccharomyces cerevisiae. Pertenece a la familia de transportadores abc (atp binding cassette) que incluye prote\u00ednas humanas como cftr (cystic fibrosis conductance transmembrane regulator) y mrp1 (multidrug resistance-associated protein).  el objetivo de este trabajo ha sido profundizar en el estudio de las relaciones estructurales y funcionales de la prote\u00edna ycf1p.  en una primera aproximaci\u00f3n se crearon mediante mutag\u00e9nesis dirigida veintid\u00f3s mutaciones \u00fanicas en residuos conservados de los dominios de uni\u00f3n a nucle\u00f3tidos (nbd1 y nbd2), transmembrana (tmd2) y regulador (r) de la prote\u00edna.  la caracterizaci\u00f3n de los mutantes ha llevado a la identificaci\u00f3n de residuos esenciales para la biog\u00e9nesis (ile711, leu712, phe713, glu927 y gly1413), para la funci\u00f3n (gly663, gly756, asp777, gly1306 y gly1311) y para la regulaci\u00f3n (ile840, tyr855, ala910 y arg1143) de ycf1p. Una segunda aproximaci\u00f3n ha consistido en el an\u00e1lisis de supresi\u00f3n intrag\u00e9nica de cinco mutaciones localizadas en los dominios nbd1 (g663v, g756d y d777n) o nbd2 (g1306e y g1311r) e implicadas en la uni\u00f3n y\/o hidr\u00f3lisis de atp. Todos los revertientes de los mutantes g663v, g756d, g1306e y g1311r resultaron ser reversiones de la mutaci\u00f3n primaria a la secuencia silvestre.  en cambio se han aislado trece mutaciones supresoras de la mutaci\u00f3n d777n localizadas en los dominios: tmd1 (v5431i y f565l), tmd2 (a1003v, a1021t, a1021v, n1027d, q1107r, g1207d, s1212l y w1225c), nbd1 (s674l) y nbd2 (r1415g). Esta localizaci\u00f3n demuestra la interacci\u00f3n f\u00edsica y\/o funcional de nbd1 con los otros tres dominios.La caracterizaci\u00f3n de los mutantes supresores indica que el mecanismo de supresi\u00f3n implica una alteraci\u00f3n en la especificidad de sustrato.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>An\u00e1lisis de dominios funcionales del transportador abc, ycf1p, mediante supresi\u00f3n intragenica y mutagenesis dirigida.<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 An\u00e1lisis de dominios funcionales del transportador abc, ycf1p, mediante supresi\u00f3n intragenica y mutagenesis dirigida. <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Juan  Manuel Falcon Perez <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Aut\u00f3noma de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 30\/11\/1999<\/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>Pilar Eraso Mazmela<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: alonso Rodriguez navarro <\/li>\n<li>Juan  Jos\u00e9 Arag\u00f3n reyes (vocal)<\/li>\n<li>Jes\u00fas Solera Garc\u00eda (vocal)<\/li>\n<li>Juan a Mar\u00eda Sempere condere (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Juan Manuel Falcon Perez La prote\u00edna vacuolar ycf1p (yeast cadmium factor) est\u00e1 implicada en la resistencia a 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