Duration
20h Th, 20h AUTR
Number of credits
Lecturer
Christian Damblon, Mireille Dumoulin, André Matagne, N..., Damien Sluysmans, Marylène Vandevenne
Coordinator
Language(s) of instruction
French language
Organisation and examination
Teaching in the second semester
Schedule
Units courses prerequisite and corequisite
Prerequisite or corequisite units are presented within each program
Learning unit contents
1. Aim of the module
This module presents a range of analytical and biophysical methods used to characterize proteins. The techniques covered include optical spectroscopy-UV-visible absorption, linear and circular dichroism, fluorescence, and infrared and Raman vibrational spectroscopy-static and dynamic light scattering (SLS and DLS), and thermodynamic methods such as differential scanning calorimetry (DSC), isothermal titration calorimetry (ITC), and differential scanning fluorimetry (DSF).
The module also introduces methods for the real-time analysis of biomolecular interactions, including surface plasmon resonance (SPR), biolayer interferometry (BLI), and quartz crystal microbalance (QCM), as well as three single-molecule approaches: FRET, atomic force microscopy (AFM), and optical tweezers.
For each method, the fundamental principles, technical aspects, and main applications to protein research are presented. These topics are explored further through sessions devoted to the analysis and presentation of scientific articles and through practical demonstration sessions held in the afternoons.
The selected articles focus primarily on protein stability and folding, that is, the process by which proteins acquire their three-dimensional structure. Several model systems, including ß-lactamases, single-domain antibody fragments, and lysozymes, are examined in detail using the theoretical concepts covered in the course and data from the scientific literature.
2. Course content: lectures
2.1. Optical spectroscopy applied to the study of biological macromolecules: UV-visible absorption, infrared and Raman spectroscopy, linear and circular dichroism, and fluorescence. 9h -- Christian Damblon.
2.2. Static and dynamic light scattering (SLS and DLS) applied to the analysis of biological macromolecules. 2h -- Marylène Vandevenne.
2.3. Thermodynamic methods applied to the analysis of biological macromolecules and their interactions: isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), and differential scanning fluorimetry (DSF). 2h -- Marylène Vandevenne.
2.4. Methods for the real-time analysis of biomolecular interactions: SPR, BLI, and QCM. 2h -- Mireille Dumoulin.
2.5. Single-molecule approaches: AFM, optical tweezers, and FRET. 3h -- Damien Sluysmans and Christian Damblon.
3. Course content: tutorials and practical sessions
The use of biophysical methods to study protein folding and stability is illustrated through the analysis and presentation of scientific articles (André Matagne).
Practical demonstrations are performed using the instruments available at the CIP (André Matagne, Marylène Vandevenne, and Romain Malempré).
Attendance at the demonstration sessions is mandatory.
Learning outcomes of the learning unit
By the end of this course unit, students will have acquired an overview of the main analytical tools available for characterising proteins at the molecular level. Particular attention will be given to the biophysical methods used to investigate protein stability, folding, and conformational dynamics and, more generally, those of biological macromolecules.
Students will be able to select the most relevant approaches according to the properties of the protein sample and the objectives of the analysis. They will understand the importance of these methods for basic biochemical characterisation and the preparation of more advanced structural studies, as well as for the quality control of samples intended, in particular, for biomedical applications.
Prerequisite knowledge and skills
Basic knowledge of chemistry and physics, together with a fundamental understanding of the structure of biological macromolecules.
Planned learning activities and teaching methods
Teaching combines lectures delivered using PowerPoint presentations and explanations at the blackboard, sessions devoted to the analysis and presentation of scientific articles, and practical demonstrations performed using the instruments available at the CIP.
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
Platform(s) used for course materials:
- MyULiège
Further information:
PowerPoint presentations and/or lecture notes will be made available to students no later than the end of the corresponding teaching sessions. These materials are intended to facilitate the study of the course content. Unless otherwise indicated, their use is recommended but not compulsory for achieving the intended learning outcomes of this course unit.
The scientific articles to be studied will be provided progressively as the course proceeds.
Recommanded textbooks :
- Biophysical Chemistry, C.R. Cantor and P.R. Schimmel.
- Principles of Physical Biochemistry, K.E. Van Holde, W.C. Johnson, P.S. Ho.
- Biological Spectroscopy, I. D. Campbell and R.A. Dwek.
- Physical Biochemistry: Principles and Applications, D. Sheehan.
- The Physical and Chemical Basis of Molecular Bioloy, Thomas E. Creighton.
- The Biophysical Chemistry of Nucleic Acids and Proteins, Thomas E. Creighton.
Exam(s) in session
Any session
- In-person
oral exam
Further information:
Assessment consists of an oral examination and a written examination.
The oral examination is conducted in the presence of the module coordinator, André Matagne, and the main lecturer responsible for the theoretical courses, Christian Damblon. The participation of the other lecturers, Mireille Dumoulin, Damien Sluysmans and Marylène Vandevenne, is optional and may vary from one examination session to another.
The oral examination covers the material taught by the lecturers who are present during the examination. It adopts an integrated approach aimed at assessing the student's ability to mobilise their knowledge, establish links between the different methods studied, and select the most appropriate tools to characterise a protein or, where relevant, another biological macromolecule. The discussion focuses in particular on the scientific articles analysed during the course, as well as on additional articles that students may have been asked to read.
A written examination covering the material taught by lecturers who do not take part in the oral examination is also organised. It accounts for no more than 30% of the final grade. The exact weighting is determined by the teaching staff and communicated to the students before the assessment period.
The written examination may take place before or after the oral examination, according to arrangements communicated to the students before the assessment period. At the end of the first examination, students are de facto informed of the parts of the course material that will be assessed in the second. As the oral and written examinations constitute two complementary components of the same assessment, participation in both is compulsory. Participation in the first examination is a prerequisite for access to the second. Any absence is dealt with in accordance with the applicable regulations.
Assessment takes into account the student's mastery of the fundamental principles underlying the methods studied, their understanding of the possibilities and limitations of these methods, their ability to select and combine appropriate approaches, their capacity to critically analyse experimental data and scientific articles, and the rigour and clarity of their answers.
Active, in-person participation in all demonstration sessions is compulsory. In accordance with Article 61 of the General Regulations for Studies and Examinations, students who fail to meet this requirement may not take the assessments organised as part of this course unit and may not obtain the corresponding credits.
Work placement(s)
Non applicable
Organisational remarks and main changes to the course
Classes are held according to the schedule communicated to the students, unless otherwise indicated. Any change to the schedule, including the cancellation or rescheduling of a class, will be communicated to the students by the lecturer, either directly during a class or by email, where appropriate through the class representative.
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 B6c, (room 3/1), 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