Proteome. Electrophoresis. 2D Electrophoresis. Multidimensional chromatography. Protein Mass Spectrometry. Protein identification. Protein sequencing methods. Differential proteomics methods. Protein covalent modifications and their analysis. Differential and functional proteomics. In vitro and in vivo analysis for protein complexes.
Text book:
Biochimica e biologia molecolare. Principi e tecniche. di K. Wilson and J. Walker. Raffaello Cortina editore
Reviews and scientific articles dealing with main topics
Learning Objectives
Knolewdge acquired:
Preparation of samples, extraction, purification. Electrophoresis, chromatography and mass spectrometry: principles and advanced techniques. Proteomics techniques and approach. Differential and functional proteomics.
Competence acquired
Lessons have the aim to show the student the right methodology for the analysis of proteome and for the identification of proteins. The analysis of the obtained results is also discussed. Theoretical and practical aspects of a proteomic study.
Skills acquired (at the end of the course):
Students will be able to set up experiments with a proteomic approach.
Prerequisites
Courses to be used as requirements (required and/or recommended)
Lectures (hours): 40
Laboratory (hours): 12
Slides of lessons will be downloaded on Moodle.unifi. The required password will be provided to the students during lessons.
Further information
It is strongly recommended to attend the lectures and lab activities
Teaching tools:
The instrumentation of the Biochemistry section of the Department of Experimental and Clinical Biomedical Sciences, will be used to perform demonstrative lessons and practice.
Office hours: every day by appointment.
Type of Assessment
The student will be examinated on the topics discussed during the course e during the lab activity. In particular the knowledge, the deepening, the reasoning skills, the overall view of the Proteomics, the clarity of the exposition, the use of a language with appropriate terminology will be considered.
Course program
Proteome. Strategies for proteomic studies. Sample preparation. Bidimensional Electrophoresis. In gel protein staining. Western Blot. Protein Image analysis. HPLC. Protein Mass Spectrometry: ionization souces and analysers. In-gel digestion and finger print. Protein identification. Protein sequencing methods. LC-MS coupling. LC-MS/MS. Quantitative analysis of proteins. Differential proteomics methods (DIGE; ICAT; iTraq.). SILAC. Protein covalent modifications and their analysis. Differential and functional proteomics. Protein arrays. In vitro and in vivo analysis for protein complexes. Interaction sites identification and characterization.