Methods in biochemical investigation

A.A. 2021/2022
Crediti massimi
Ore totali
Obiettivi formativi
The study of proteins through protein biochemistry encompasses many fields of biology, including structural and molecular biology, enzymology, pharmaceutical research and development, food science, plant biology, and more. The course aims to provide students with a thorough overview (theoretical and practical) of the most important and recent methods applied to enzymology (focusing on the central theme of enzymes) and to structural biology (i.e. X-ray crystallography, single-molecule Cryo-electron microscopy), focusing in particular on the scientific questions that such techniques can help answer.
Risultati apprendimento attesi
By the end of the course, students will have acquired a good understanding of the experimental strategies applied in biological chemistry, in particular regarding protein enzymology and structural biology. Moreover, students will have acquired the ability to correlate the theoretical and experimental connections among the taught disciplines and understand their relevance to biochemical developments.
Programma e organizzazione didattica

Edizione unica

Secondo semestre
In relazione alle modalità di erogazione delle attività formative per l'a.a. 2021/22, verranno date indicazioni più specifiche nei prossimi mesi, in base all'evoluzione della situazione sanitaria.
The program of the Advanced Enzymology part of the course includes the following main topics: enzyme reaction mechanisms and related rate equations under steady-state conditions; one- and two-substrate enzyme reactions; reversible enzyme reactions; techniques for the study of steady-state phase of enzyme reactions; enzyme assays; determination of enzyme kinetic parameters; enzyme inhibition; experimental design and interpretation of results in the kinetic study of enzyme reactions.
The Structural Biology program is focused on the 3D structure determination and analysis of proteins by using biocrystallographic and bioinformatic techniques. Experimental techniques for protein crystal growth will be described in detailswith application to soluble and membrane proteins. X-ray diffraction on protein crystals will be treated extensively (X-ray sources, synchrotron light, atomic scattering factors, structure factors, the "phase problem", phasing by molecular replacement and heavy atoms, crystallographic refinement). Protein model validation techniques will be analyzed, together with the main features of the Protein Data Bank. In addition, the theoretical basis and the practical applications of single-molecule cryo-EM techniques will be treated.
A revision of the topics covered by the basic biochemistry and molecular biology classes included in the bachelor curriculum before attending the course is highly recommended.
Metodi didattici
Teaching Mode: Classroom lectures supported by projected material with common discussions on experimental design, data analysis, and specific case studies. Demonstration of experiments on enzyme kinetics will be provided using specific computer simulation programs.
Materiale di riferimento
Students can refer to their text of Biochemistry for the general aspects of the structure of enzymes and catalysis. Further useful insights can be found in:
- N. Price and L. Stevens, "Fundamentals of Enzymology - Cell and Molecular Biology of Catalytic Proteins" 3rd Ed., Oxford University Press, 1999.
- A. Fersht, "Structure and Mechanism in Protein Science. A Guide to Enzyme Catalysis and Protein Folding", WH Freeman & Co., 1999
For the part of Structural Biology, the following textbooks are suggested:
- Ducruix and R. Giegé (Oxford University Press)
- Crystallography made crystal clear. A guide for users of macromolecular models. By Gale Rhodes (Elsevier)
Copies of the slides projected in the classroom as well as other materials will be made available through the course website on the ARIEL platform of the University of Milano ( By no means, this material replaces the lectures or a textbook. The material is made available only to registered students of the Degree Course in Molecular Biotechnology and Bioinformatics and should not be distributed to others.
Modalità di verifica dell’apprendimento e criteri di valutazione
The evaluation of the student's performance will be based on a written examination with 3-4 open-answer questions and/or exercises both for the enzymology (50% of the final score) and for the structural biology (50% of the final score) parts. Open questions are designed to evaluate the ability of the student to concisely and critically describe the major topics covered in the class. Exercises are meant to evaluate the ability of the student to solve applied problems. Each student is allowed to give the exam on the enzymology part of the program in oral form, by informing the teacher in advance.
The final grade will result from the joint evaluation of each candidate by the two teachers.
Lezioni: 48 ore
Ven 08:30-12:30 (previo appuntamento via e-mail)
Edifici Biologici, via Celoria 26 - torre C - piano 5
Giovedi, ore 10:30-12:30
Dip. di Bioscienze, Torre C, 5 piano