Development of probes for molecular imaging of metallo-carboxypeptidases: Synthesis of FRET substrates, activable “smart probes”, phosphinic pseudopeptides on modular/powdered solid supports using the Fmoc-based chemistry (solid-phase peptide synthesis, SPPS). Synthesis of small libraries of phosphinic pseudopeptides on modular solid supports. Purification of small organic compounds by RP-HPLC. Analysis of synthesized compounds by ESI-MS. Functional-Structural characterization of FRET substrates, activable “smart probes” and phosphinic pseudopeptides towards target metallo-carboxypeptidases with biomedical applications (ie., TAFI, CPA4, among others).
Optimization of high-level expression in Pichia pastoris of carboxypeptidases and carboxypeptidase inhibitors with biomedical applications: development and process scale production of recombinant proteins in the early stages of cloning, expression analysis in Erlenmeyer flask-grown cultures, fermentation at pilot plant scale using Pichia pastoris expression system, as well as fermentation with high cell density cultivation (HCDC) strategy.
Doctoral thesis: Kinetic and proteomic identification of protease inhibitors in marine invertebrates. Characterization of a carboxypeptidase inhibitor isolated from the mollusk Nerita versicolor. (defense date: October 23 2012) (http://hdl.handle.net/10803/117662)
Covaleda. G., Alonso del Rivero, M., Chávez, M.A., Avilés, F.X., and Reverter, D. Crystal Structure of Novel Metallocarboxypeptidase Inhibitor from Marine Mollusk Nerita versicolor in Complex with Human Carboxypeptidase A4. J Biol Chem. 2012; 287(12): 9250-9258.
Fraga, H., Graña-Montes, R., Illa, R., Covaleda, G., and Ventura, S. Association between foldability and aggregation propensity in small disulfide-rich proteins. Antioxidants & Redox Signaling. 2014; 21(3): 368-383.
Bronsoms, S., Covaleda, G., and Avilés, F.X. Intensity Fading MALDI-TOF MS en proteómica functional e interactómica. Manual de Proteómica. Spanish Proteomics Society SEProt. Chapter 31. 2014; 695-708.
Fernández, D., Pallarès, I., Covaleda, G., Avilés, F.X., and Vendrell, J. Metallocarboxypeptidases and their inhibitors: recent developments in biomedically relevant protein and organic ligands. Current Medicinal Chemistry. 2013; 20(12): 1595-1608.