Synthesis and optimization of new sphingolipid sensors for metabolism and trafficking studies

• Autores: Ana Pou Cabello
• Directores de la Tesis: Antonio Delgado Cirilo (dir. tes.), José Luis Abad Saiz (dir. tes.)
• Lectura: En la Universitat de Barcelona ( España ) en 2017
• Idioma: español
• Tribunal Calificador de la Tesis: Albert Moyano Baldoire (presid.), Maria del Pilar Marco Colas (secret.), Miriam Royo Expósito (voc.)
• Programa de doctorado: Programa de Doctorado en Química Orgánica por la Universidad de Barcelona
• Materias:
  • Química
    • Bioquímica
      • Lípidos
      • Procesos metabólicos
      • Biología molecular
• Enlaces
  • Tesis en acceso abierto en:  Dipòsit Digital de la UB  TDX 
• Resumen

  • Over the past decade, sphingolipids have emerged as a new class of modulators of various cell functions. Ceramide, which is the central molecule in the biosynthesis of sphingolipids and glycosphingolipids, is involved in the regulation of different cellular events, including cell senescence, differentiation, and apoptosis. In the de novo pathway, ceramide is biosynthesized from L-serine in four steps, the last one being the Δ4-desaturation of dihydroceramide to ceramide by the action of dihydroceramide desaturase (Des1).

    The discovery of new Des1 inhibitors with improved potency and selectivity would be greatly accelerated with the help of high-throughput screening (HTS) assays. Toward this end, the design of an efficient probe to quantify the enzyme activity is capital. In this Thesis, we wish to report that (Z) Δ6 dihydroceramide is a suitable Des1 substrate and that the resulting Δ4,6-diene product can be efficiently trapped in a Diels Alder reaction with a fluorescent TAD derivative for further quantification. In addition, the unanticipated observation that the isomeric (E) Δ6 dihydroceramide is a non competitive Des1 inhibitor provides valuable information about the conformational requirements of the enzyme active site.

    The design of HTS assay to monitor Des1 activity on solid support by anchoring the (Z) Δ6 dihydroceramide as Des1 substrate and the use of a fluorescent reporter in an array system will be presented.