Proteomics and protein characterization


E. Biasini, G. Lolli, C. Del Bianco, R. Belli , D. Peroni (University of Trento)

R. Fiala, K. Kubichek, P. Kaderavek, J. Novacek (CEITEC Masaryk University)

The aim of the course is to provide a broad knowledge of methods and applications within Proteomics, exploring advanced methods for protein characterization and functional proteomics. Emphasis will be placed on purification and separation methods, on MS-based strategies and on high-end proteomics workflows and technologies, including NMR, Crystallography and Cryo-EM.  Moreover, specific examples of practical applications and case studies will be provided.

Upon successful completion of this course, participants should:

  • Understand the principles underlying the expression, purification and characterization of proteins.
  • Discuss the importance of using mass spectrometry in the study of proteins in complex biological samples, explaining their potential and limitations.
  • Critically choose the proteomic approach for the qualitative and quantitative analysis that is best suited to answer the scientific question of interest.
  • Evaluate the appropriate method for sample preparation considering compatibility with mass spectrometry analyses, as well as the purpose of the experiment.
  • Understand the principle of operation of the main components of the mass spectrometer, optimizing the parameters that determine its performance.
  • Read mass spectra (e.g., de novo sequencing) and understand the logic of complex dataset analyses via software.
  • Consult online databases and use the most common software for processing experimental data.
  • Interpret, process and present experimental results.
  • Understand the principles behind Nuclear Magnetic Resonance (NMR) spectroscopy, X-ray crystallography, and cryoelectron microscopy (CryoEM) for the structural analysis of proteins and protein complexes.
  • Read and understand scientific articles in the field of protein analysis.