PROTEIN ENGINEERING
- Overview
- Assessment methods
- Learning objectives
- Contents
- Full programme
- Delivery method
- Teaching methods
- Contacts/Info
The student who attends this course will be asked to apply the knowledge acquired in the core curriculum of the First Cycle Degree: Biochemistry (in particular solid knowledge regard-ing the structure and properties of proteins are required); Molecular Biology, (recombinant DNA techniques - manipulation, amplification, cloning - but also gene expression regulation mechanisms); Microbiology (distinctive features of the microorganisms used as heterologous hosts for the production of recombinant proteins and their maintenance in culture). The knowledge and practical skills acquired during the course of Biochemical Methodologies are also required.
Evaluation of learning will take place through a written test aimed at assessing the learning achievements and therefore the understanding the problems discussed during classes and the ac-quired knowledge related to the "Protein Engineering". The student will answer to 4 open ques-tions (each evaluated up to 8 points).
The outcome of the exam will be out of thirty: the exam is considered passed with a mark of at least 18/30.
The criteria according to which the acquired knowledge and skills will be assessed are:
1. the degree of depth of the subject matter of the question;
2. the critical ability to rework and link the knowledge acquired regarding the theoretical bases of the structure-function relationship in proteins;
3. the ability to apply the knowledge acquired to develop a protein engineering project and to de-sign, based on the characteristics of a specific protein of interest, an expression strategy in one or more suitable heterologous systems;
4. the ability to propose solutions to specific problems by using the tools and strategies discussed during teaching;
5. the clarity of the exposed concepts and the use of appropriate scientific terminology.
This course of Protein Engineering is fundamental to the preparation of a student in BBHI. The course aims to provide students with a solid preparation in the design and production of recombinant proteins, to be used as biotechnological tools in the biomedical (as therapeutic agents) and industrial (as components of bioconversion and biocatalysis production process-es), in line with the educational objectives proposed by the Degree.
In particular, the course aims to provide the knowledge of the different techniques in the bi-otechnological application of protein engineering (ex novo design of protein structures, ra-tional design of new functions, molecular modeling and docking, site-directed and random mutagenesis, gene recombination), as well as methods through which it is possible to isolate the optimized protein variants in a given property.
Students will be encouraged to evaluate the advantages and limitations of the various ap-proaches used to modify proteins properties. Case studies will be also discussed. Students are expected to acquire decision-making skills in a "protein engineering" project, from the design phase to the production of recombinant protein variants one, as well as to be able to face and solve the various problems related to the process.
At the end of the course, students will be able to:
• explain at the molecular level the fundamental elements of the structure-function relation-ship of proteins;
• critically apply the acquired notions (theoretical and experimental) to the engineering of the functionality and stability of a protein;
• evaluate, extract and synthesize the information acquired, when relevant for a specific pro-ject or case study;
• communicate the notions learned effectively both orally and in writing using appropriate scientific language.
In conclusion, students will acquire decision-making skills in a "protein engineering" project, from the design phase to the production of the recombinant protein variants, being able to face and solve the various problems related to the process.
Lecturer: Prof. Loredano Pollegioni
6 credits or 48 hours of lectures. The program is divided into two parts which deal with the following topics:
a) Structure and function relationship in proteins
- Introduction to the structure of proteins; structural classification of proteins
- Protein synthesis: mechanisms and regulation
- Experimental and in silico methods of protein structure determination
- Post-translational changes and protein targeting
- Protein folding (in vitro and in vivo)
- Protein degradation (lysosomal and proteasomic system)
- Membrane proteins: relationship between structure and function
b) Protein Engineering:
- De novo protein design (of structures and functions): methods and examples
- Rational design: theory and experimental methods. Examples
- Molecular docking: theory and examples
- Irrational design: theory and experimental methods of directed evolution. Examples
- Screening methods
- Protein stability engineering: theory, methods and examples.
- Metabolic engineering and systems biology: integration.
The program is divided into two parts which deal with the following topics:
a) Structure and function relationship in proteins
- Introduction to the structure of proteins; structural classification of proteins
- Protein synthesis: mechanisms and regulation
- Experimental and in silico methods of protein structure determination
- Post-translational changes and protein targeting
- Protein folding (in vitro and in vivo)
- Protein degradation (lysosomal and proteasomic system)
- Membrane proteins: relationship between structure and function
b) Protein Engineering:
- De novo protein design (of structures and functions): methods and examples
- Rational design: theory and experimental methods. Examples
- Molecular docking: theory and examples
- Irrational design: theory and experimental methods of directed evolution. Examples
- Screening methods
- Protein stability engineering: theory, methods and examples.
- Metabolic engineering and systems biology: integration
The "Protein Engineering" course consists of 48 hours of lectures. All activities will take place in the presence of the instructor in charge. Each lesson will be carried out by treating a specific topic with the aid of powerpoint presentations, available in advance on the e-learning platform. During the lessons, case studies published in scientific jour-nals of the field will also be presented, which will be provided as didactic material.
Reception preferably by appointment (by request via e-mail). The lecturer replies only to e-mails signed and from the @ uninsubria.it domain. We do not respond to requests for confirmation of information already available - for example, request of the exam date.
Prof. Pollegioni is available for in-depth meetings or discussions for groups of students.
Slides of the lessons: downloadable from the e-learning site
Selected books:
Arnold F. H., Georgiou G. Directed Enzyme Evolution: Screening and Selection Methods (Methods in Molecular Biology) (Humana Press)
Arnold F. H., Georgiou G. Directed Evolution Library Creation: Methods and Protocols (Methods in Molecular Biology) (Humana Press)
Branden C. e Tooze J., Introduction to protein structure (Garland Publ.)
Bross P. e Gregersen N. Protein misfolding and disease, Methods in Molecular Biology, vol. 232 (Humana Press).
Garett-Grisham "Principi di Biochimica" (PICCIN)
Matthews, Van Holde, Ahern, “Biochimica” (Casa Editrice Ambrosiana)
Voet D, Voet JG, Pratt CW "Fondamenti di Biochimica" (Zanichelli)
The material useful to complete the preparation (scientific papers and reviews) will be downloadable from the e-learning site. Additional material will also be available at request.