Per citar aquest document: http://ddd.uab.cat/record/101079
Study of molecular mechanisms in glycoside hydrolases and transferases by ab initio molecular dinyamics / Albert Ardèvol Grau ; direcció de Carme Rovira Virgili ; tutorial del Dr. Xavier Parés Casasampera
Ardèvol Grau, Albert
Rovira i Virgili, Carme, dir. (Institució Catalana de Recerca i Estudis Avançats)
Parés i Casasampera, Xavier, dir. (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Universitat Autònoma de Barcelona. Departament de Bioquímica i Biologia Molecular

Publicació: [Barcelona] : Universitat Autònoma de Barcelona. Departament de Bioquímica i Biologia Molecular, 2012
Descripció: 1 recurs electrònic (182 p.)
Resum: Carbohydrates had historically been associated to two biological functions: energy storage and structural support. However, in the last decades, new complex structures of oligosaccharides have been found to play vital roles in many biological processes, such as signal transduction, immune response, cell differentiation and cancer development, among others. Advances in the functional understanding of carbohydrate-protein interactions represented a breakthrough in the field of glycobiology and glycochemistry, opening a new branch of potential therapeutic targets (carbohydrate acting enzymes), glycomimetic drugs and biomarkers. The bottleneck in the field of glycochemistry is the synthesis of complex saccharides; hence many efforts have been devoted to the development of novel enzymatic strategies for carbohydrate synthesis. Glycoside transferases (GT) and glycoside hydrolases (GH) are the enzymes that catalyze the formation and the cleavage of the glycosidic linkage respectively. They are used in complex oligosaccharides synthesis, and recently they have been engineered to produce enzymes with particular substrate specificities or even activities. In spite of these advances, the understanding of the molecular mechanisms of enzymatic carbohydrate synthesis and degradation is far from complete. Structural studies have shown that the puckering of the sugar ring at the cleavage point must change during catalysis. Knowing the conformational catalytic itinerary has an impact in the design of GHs inhibitors. However, these itineraries are not known for all families of GHs. On the other hand, the saccharide puckering is not an issue in GTs, but the reaction mechanism is not known. In fact, the glycosidic bond formation in GTs remains one of the most intriguing and unanswered questions in the field of glycobiology. The coming of age of powerful theoretical methods such as quantum mechanics / molecular mechanics (QM/MM) and ab initio molecular dynamics (AIMD) has enabled the elucidation of complex reactive processes in proteins and enzymes. In particular, the modeling of the Michaelis complex and the reaction mechanisms of GHs highlighted the interplay between electronic and structural changes that preactivate the substrate for catalysis. Some of these changes can already be anticipated by analyzing the conformational energy landscape of the substrate. Part of the research of this Thesis complements previous studies of our group by analyzing the factors that govern substrate distortion in GHs. In this respect, it extends the use of conformational free energy landscapes of simple sugars to predict the conformation of the substrate in Michalis complexes. Additionally, the molecular mechanism of retaining glycoside transferases is elucidated. This Thesis is organized as follows: Chapter I contains an introduction of the enzymes studied (GHs and GTs) and presents the main objectives of this work. The theoretical methods used are detailed in Chapter II. Chapters III to V are focused on enzyme-substrate interactions affecting the conformation of the substrate in GHs. Concretely; in Chapter III we test how mutation of the acid/base catalytic residue, the use of a substrate-like thio-analogue inhibitor or fluorometric aglycons affects the distortion of the substrate. In Chapter IV we study the influence of the enzyme-substrate interactions through the 2-OH, in particular the effect of the commonly used 2-deoxy-2-fluoro substitution. The conformational itinerary of this inhibitor during catalysis is modeled in Chapter V. In Chapter VI, the conformational flexibility of β-D-mannopyranose and α-L-fucopyranose molecules is investigated. The topologies of their corresponding conformational free energy landscapes are related with the observed crystallographic structures of β-mannosidases and α-fucosidases, and the predictive potential of such calculations is discussed. Chapter VII focuses on trehalose 6-phosphate synthase (a family 20 retaining GT that belongs to fold type B). The mechanism of glycosidic bond formation in this enzyme is elucidated. Finally, in Chapter VI, the main conclusions of this work are summarized.
Nota: Tesi doctoral - Universitat Autònoma de Barcelona. Departament de Bioquímica i Biologia Molecular, 2012
Nota: Descripció del recurs: el 01 setembre 2012
Drets: ADVERTIMENT. L'accés als continguts d'aquesta tesi doctoral i la seva utilització ha de respectar els drets de la persona autora. Pot ser utilitzada per a consulta o estudi personal, així com en activitats o materials d'investigació i docència en els termes establerts a l'art. 32 del Text Refós de la Llei de Propietat Intel·lectual (RDL 1/1996). Per altres utilitzacions es requereix l'autorització prèvia i expressa de la persona autora. En qualsevol cas, en la utilització dels seus continguts caldrà indicar de forma clara el nom i cognoms de la persona autora i el títol de la tesi doctoral. No s'autoritza la seva reproducció o altres formes d'explotació efectuades amb finalitats de lucre ni la seva comunicació pública des d'un lloc aliè al servei TDX. Tampoc s'autoritza la presentació del seu contingut en una finestra o marc aliè a TDX (framing). Aquesta reserva de drets afecta tant als continguts de la tesi com als seus resums i índexs.
Llengua: Anglès.
Document: Tesis i dissertacions electròniques ; doctoralThesis
Matèria: Glicòsids ; Transferases ; Dinàmica molecular

Adreça alternativa: http://hdl.handle.net/10803/83945


182 p, 3.3 MB

El registre apareix a les col·leccions:
Documents de recerca > Tesis doctorals

 Registre creat el 2012-11-23, darrera modificació el 2016-04-15



   Favorit i Compartir