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ESTUDIS DE A11 ACIL-Coa DESSATURASES IMPLICADES EN

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ESTUDIS DE A11 ACIL-Coa DESSATURASES IMPLICADES EN ESTUDIS DE '11 ACIL-CoA DESSATURASES IMPLICADES EN LA BIOSÍNTESI DE FEROMONES SEXUALS DE LEPIDÒPTERS Montserrat Serra Mas Barcelona, 2006 DEPARTAMENT DE BIOQUÍMICA I BIOLOGIA MOLECULAR FACULTAT DE BIOLOGIA Programa de Doctorat de Biotecnologia Bienni 2002/2004 ESTUDIS DE '11-ACIL-CoA DESSATURASES IMPLICADES EN LA BIOSÍNTESI DE FEROMONES SEXUALS DE LEPIDÒPTERS Memòria presentada per Montserrat Serra Mas per optar al títol de Doctor per la Universitat de Barcelona Treball realitzat en el Departament de Química Orgànica Biològica de l’Institut d’Investigacions Químiques i Ambientals de Barcelona (C.S.I.C.) Directors Tutor Jordi Bujons Vilàs Montserrat Busquets Abió Científic titular Professora titular Dpt. Química Orgànica Biològica Dpt. Bioquímica y Biologia Molecular Institut d’Investigacions Químiques Facultat de Biologia i Ambientals de Barcelona Universitat de Barcelona (I.I.Q.A.B.-C.S.I.C.) Gemma Fabriàs Domingo Investigador científic Dpt. Química Orgànica Biològica Institut d’Investigacions Químiques i Ambientals de Barcelona (I.I.Q.A.B.-C.S.I.C.) What we know is a drop. What we don't know is an ocean. Isaac Newton (1643-1727) Como siempre: lo urgente no deja tiempo para lo Importante. Mafalda AGRAÏMENTS Quan s’arriba al punt de redactar els agraïments de la tesi doctoral vol dir que ja queda poca cosa per fer. A més, aquest és un apartat especialment important ja que saps del cert que aquestes dues pàgines les llegirà tothom. En aquest moment mires enrera i recordes el dia en el que vas entrar per primera vegada en un laboratori. Tenia claríssim que la meva especialitat havia d’ésser la bioquímica i els meus primers passos els vaig donar en un laboratori de la setena planta de la facultat de Química. Encara sembla que fos ahir quan una nena espantada i extremadament tímida s’estava asseguda en un escriptori llegint una tesina sobre la malat deshidrogenasa citosòlica de porc. Han passat ja vuit anys des d’allò i ara toca donar les gràcies a tota la gent que ha fet possible que aquesta tesi existeixi. Primer de tot, i com ha de ser en qualsevol tesi voldria agrair la direcció d’aquesta tesi als Drs. Gemma Fabriàs i Jordi Bujons, per les estones dedicades a corregir coses i a proposar noves línies de treball. També a la Dra. Montserrat Busquets per haver acceptat de ser la tutora d’aquesta tesi. També he d’agrair al Dr. Francesc Camps que en el seu dia confiés en mi per realitzar aquesta tesi. Així com a tots els integrants del Profero: Jesús Joglar, Pere Clapés, Conxita Solans.... Una menció a part mereix el Dr. Benjamí Piña. Sense les teves instruccions, els teus consells i les estones que has passat amb la teva becària “externa” aquesta tesi no sé pas si s’hauria arribat a acabar mai. Recordo ara totes les hores passades comentant si uns llevats amb '11 de T. ni creixien amb C10 o si la seqüència de la processionària estava bé o no. Moltíssimes gràcies.... Qui també sempre ha estat allà ha estat el Dr. José Luis Abad (JLAS per als amics o almenys aquí al CSIC) sempre obsequiant-nos amb les seves interessants converses. Molta sort amb les opos!!! Ànims que tu pots!! M’agrada tenir un record per al meu predecessor en el tema de la Biologia Molecular de les dessaturases, el Dr. Sergio Rodríguez. De tu Sergi, vaig aprendre moltíssimes coses (i no només on era el material que hauria de fer servir durant la tesi!!!). Gràcies a tu i a la Dra. Gemma Villorbina em vaig posar al dia del tema de les dessaturases i els insectes dels que no en tenia ni idea (a part de l’acollida al teu piset). i D’aquella primera època també vull recordar la resta de gent que ens movíem pels laboratoris 213, 004 i 314: Carme, Marta, Lidia Batlle, Lidia Roura, Ismael, Cristina, Òscar, Itziar, Jorge i Tània. No em deixo en Carles Soldevila del que mai no sabies si estava aquí o a la lluna de València ni tampoc la Victòria que definiria una miqueta com la meva psicòloga particular ja que sempre em deixa parlar i m’escolta les meves paranoies. Vull també expressar el meu agraïment a la gent de l’Institut de Biologia Molecular de Barcelona per haver-me deixat utilitzar els aparells i els kits que m’han fet falta en moments molt puntuals d’aquest treball i en especial la Sílvia. Una abraçada molt forta a la gent del laboratori 516: l’Alba (últimament sempre et trobo a l’hora de dinar (quan vaig mooolt tard)), la Sue (gràcies pel protocol del Western), la Marta, la Tània (quines tertúlies a primera hora del matí), el David i la Laia (vam començar gairebé juntes i ara ja falta una miqueta menys per acabar). També he de donar les gràcies al Dr. Isidre Casals que durant aquests quatre anys de tesi ha estat el meu taxi particular. Però no només això m’ha escoltat i m’ha aconsellat sempre que ho he necessitat. I want to thank to Prof. Peter H. Buist the opportunity of working with you in Carleton University (Ottawa, Canada) and thank you for the paper. It was one of the most wonderful experiences I ever had. Thank you very much for all your help and give my best wishes for the people from the lab. I also wanted to thank the assistance given by Dr. William G. Willmore in his lab for all the microsomes work. I want also to dedicate my thought to all the people who helped me when I stayed there: Molly, John, Laura (all the best in your graduation), Kim, Amber, Amy, Nigel, Rishi, James, Ryan, Chris, Angie and Joann (I am so sorry if I didn’t write any of the names correctly). Tampoc em vull oblidar de la Dori i la Roser que sempre m’han ajudat amb el cromatògraf de gasos acoblat a l’espectròmetre de masses, quan es penjava, quan els pics no sortien prou clars....vaja! Quan hi havia problemes!!! També he d’agrair la bona feina del servei de seqüenciació del CSIC encapçalat per la Mercè Miquel. He de recordar en aquest punt als companys de fatigues en els temes de biologia molecular dins el departament: la Noemí, la Glòria, el Gerard, la Cristina i el Xavi. Cal anomenar a tots els companys amb els qui he compartit laboratori: la Mikaela, la Carme, el Pablo, l’Hèctor, el Javi Adrián (no més humiliacions al Supermanager), el Javi Ramon (m’encanta Wagner), el Dani, el Mark, la Raquel, la ii Núria, el Roger i la Begoña (amb qui hem compartit moltíssimes bones estones). També l’acollida dels Drs. Pilar Marco i Francisco Sánchez. Moltes gràcies a tota la gent de la segona planta: la Maria, l’Ariadna, el Carles, el Jordi, el Jose, la Sònia, l’Aurora i els del RUBAM per ser-hi sempre: la Isabel, la Marta, l’Anna, la Patrícia, la Meri, la Gemma, la Sílvia Saludes, la Sílvia González, la Rebeca, l’Elena, el Fabrizio, el Pedro, l’Albert, el Santi, el Dani, el Youssef, el Laurent i el Jose. I moltíssimes gràcies a l’Esmeralda, la Lourdes, el David i la Carma els millors companys que es poden tenir a l’hora de fer una tesi doctoral. Un petonàs molt fort per tots els antics enzimos: la Meri (no passis gaire fred a Munich), l’Oscar Besumbes (no maltractis molt els “nanos”), l’Oscar Alegre i la Mari, el Churri (ho sento Jose però t’ha quedat aquest nom) i la Sònia (sort a tots dos a Indianapolis), el Víctor, el Cristòbal, el Joan i el Heavy (sort amb els esterols). Dins d’aquest apartat d’agraïments també hi he de posar la meva família, els meus pares i germans que malgrat no entendre res del que explico sempre intenten preguntar com va la feina i em donen el màxim suport. Finalment, voldria agrair tot el suport i la comprensió que m’ha donat el Xavier. Ell que ha hagut d’aguantar tots els meus nervis i els meu mal humor com les coses no anaven prou bé però amb qui també he compartit totes les bones estones. Només et volia dedicar unes petites paraules: Por una mirada un mundo; por una sonrisa, un cielo; por un beso..., ¡yo no sé qué te diera por un beso! Gustavo Adolfo Bécquer (1836-1870) Moltes gràcies a tots!!!! iii ÍNDEX Índex ÍNDEX ............................................................................................................................. v ABREVIATURES ........................................................................................................... 1 NOTACIÓ DELS ÀCIDS GRASSOS........................................................................... 5 I. INTRODUCCIÓ ......................................................................................................... 9 I.1. Feromones. ......................................................................................................... 11 I.1.1. Perspectiva històrica. ................................................................................. 11 I.1.2. Les feromones en lepidòpters................................................................... 14 I.2. Dessaturases....................................................................................................... 17 I.2.1. Mecanisme catalític. ............................................................................... 20 I.2.2. '9 acil-CoA dessaturases. .......................................................................... 25 I.2.3. Citocrom b5.................................................................................................. 27 I.2.4. Dessaturases en lepidòpters. .................................................................... 28 I.3. Antecedents de la recerca sobre dessaturases en el grup de treball. ......... 30 II. OBJECTIUS .............................................................................................................. 33 III.RESULTATS I DISCUSSIÓ.................................................................................... 37 IIIa. ESTUDIS SOBRE LA '11 ACIL-CoA DESSATURASA
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