Duration
20h Th, 20h AUTR
Number of credits
Lecturer
Christian Damblon, Franck Dequiedt, Mireille Dumoulin, André Matagne, N..., Damien Sluysmans, Marylène Vandevenne
Coordinator
Language(s) of instruction
French language
Organisation and examination
Teaching in the first semester, review in January
Schedule
Units courses prerequisite and corequisite
Prerequisite or corequisite units are presented within each program
Learning unit contents
1. Aim of the module
Various analytical methods used for protein characterisation are presented. Thus, spectroscopic methods are UV-visible absorption, linear and circular dichroism, fluorescence, vibrational spectroscopy (i.e. infrared and Raman), surface plasmon resonance (SPR) and biolayer interferometry (BLI). Three 'single molecule' approaches (i.e. FRET, AFM and optical tweezers) are also studied, as well as methods for measuring interactions between molecules in real time. Classical thermodynamic approaches (i.e. DSC, ITC, DSF) are presented, as well as methods specific to the study of in vivo interactions. For each approach, the principles, technical aspects, and applications focusing on proteins will be developed, especially during the article presentation sessions and the practical work (demonstrations) that will take place in the afternoons. The selected articles focus mainly on the stability and folding (i.e. the process of acquiring the 3D structure) of proteins. A few models (e.g. beta-lactamases, single-domain antibody fragments, lysozymes) are analysed in detail, based on theoretical knowledge and data found in the literature.
2. Table of contents of theoretical courses
2.1. Optical spectroscopy applied to the study of biological macromolecules (UV-Vis absorption; infrared and Raman; linear and circular dichroism; fluorescence). 9h; Christian Damblon
2.2. Static and dynamic light scattering (SLS and DLS, respectively) for the analysis of biological macromolecules. 2h; Marylène Vandevenne
2.3. Thermodynamic methods for the analysis of biological macromolecules and the study of their interactions: a) isothermal titration calorimetry (ITC); b) differential scanning calorimetry (DSC); c) differential scanning fluorimetry (DSF). 2h; Marylène Vandevenne
2.4. Real-time biosensing methods based on e.g. SPR, BLI, QCM. 2h; Mireille Dumoulin
2.5. Single molecule approaches (AFM, optical tweezers and FRET). 3h; Damien Sluysmans & Christian Damblon
3. Tables of contents of practical courses and tutorials
The use of biophysical methods to study protein folding and stability is illustrated on the basis of data found in the litterature (André Matagne).
Demonstrations are organised using the instruments available at the CIP (André Matagne, Marylène Vandevenne, Romain Malempré).
Attendance at the demonstration sessions is mandatory.
Learning outcomes of the learning unit
Students will gain an overview of the main analytical tools available to characterise proteins at the molecular level. Particular attention will be paid to the biophysical methods available for characterising protein stability and folding, as well as for studying the conformational dynamics of biological macromolecules in general. Students should be able to select the most relevant approaches for characterising a protein sample. These methods are important both for basic biochemical characterisation, particularly with a view to more advanced structural studies, and for quality control in the preparation of samples for e.g. biomedical applications.
Prerequisite knowledge and skills
Basic chemistry and physics. Fundamental knowledge of the structure of biological macromolecules
Planned learning activities and teaching methods
Lectures are given with the help of powerpoint presentations, blackboard and chalk
Mode of delivery (face to face, distance learning, hybrid learning)
Face-to-face course
Additional information:
Face-to-face only
Course materials and recommended or required readings
Powerpoint presentations and/or lecture notes will be available at the end of the course at the latest. These materials are intended to assist students in their study; not all of them are absolutely essential for acquiring the skills required in this course and their adoption by students is not compulsory. Scientific articles will be given as they are analyzed.
Reference textbooks :
- Biophysical Chemistry, Cantor and Schimmel
- Principles of Physical Chemistry, Van Holde et al.
- Biological Spectroscopy, Campbell and Dwek
- Physical Biochemistry, Sheehan
- The Physical and Chemical Basis of Molecular Biology, Creighton
- The Biophysical Chemistry of Nucleic Acids and Proteins, Creighton
Exam(s) in session
Any session
- In-person
oral exam
Further information:
The students will be orally evaluated in the presence of the module coordinator, André Matagne, and the main lecturer for the theoretical courses, Christian Damblon. The presence of other contributors (i.e., Mireille Dumoulin, Damien Sluysmans, Marylène Vandevenne, Romain Malempré) is optional and may vary from one session to another. The oral evaluation will cover the material taught by the lecturers present at the exam, in an integrated approach that will encourage reflection and lead the student to consider the full range of tools for the complete characterization of a protein (or possibly another biological macromolecule). The discussion will notably include the series of articles studied during the course, as well as other articles recommended by the lecturers. Additionally, a written exam covering the material taught by the lecturers absent during the oral evaluation will be organized and will account for up to 30% of the final grade. This percentage will be defined by the lecturers and communicated to the students on the day of the written exam.
Active in-person participation in the demonstration sessions is mandatory. In accordance with Article 61 of the General Study and Evaluation Regulations (academic year 2024-2025), a student will not be allowed to take the assessments for this course unit or be awarded the corresponding credits if this requirement is not met.
Work placement(s)
Non applicable
Organisational remarks and main changes to the course
Lectures will be organized following the schedule known by the students. The professor will
inform the students in advance of any possible modification to this
Contacts
André Matagne, PhD, Professeur Ordinaire, Enzymologie et Repliement des Protéines, Centre
d'Ingénierie des Protéines, UR InBioS, Département des Sciences de la Vie, Institut de Chimie B6a, (bureau 1/10a), Quartier Agora, Allée du 6 Août, 13, Université de Liège, B4000 Liège (Sart-Tilman), Tel.: +32 (0)4 3663419, Email: amatagne@ulg.ac.be (best option)
Association of one or more MOOCs
There is no MOOC associated with this course.
Additional information:
Non applicable