The basic rules of protein structure must be known (biopolymers; Ba2) as well as the techniques used to purify and characterize them (analytical biochemistry ba2)

This course has as goal to give you insight in the most frequently used techniques to study proteins and proteomes, on a theoretical level on a practical level.

After this course, you should know/be able to:

-          discuss the function-relationship of proteins

-          make a good, motivated choice out of the different techniques seen, to reach a certain goal or to solve a certain problem.

-          gain insight in the different techniques to isolate and analyze protein profiles (proteomics)

-          set up a protein-purifying protocol

-          gain the ability to turn theory into practice

The theoretical aspects will be demonstrated using practica. In the first part of the course, the focus will lie on the study of proteins:

-          chemical modification of proteins: e.g. (i) chemical modification of the amino acids cysteine or lysine by vinylpiridinylation and modification by maleinic acid respectively, (ii) biotinylation of proteins

-          purification of protein mixtures with limited complexity using ion-exchange and reversed phase chromatography. The purity can be checked using SDS-PAGE

-          making a strategy for cleaving proteins

-          separation of limited complex peptide mixtures using reversed phase chromatography

-          N-terminal sequencing of proteins and peptides by Edman degradation

-          Examples of affinity-chromatography

In the following part of the course, proteomics sensu strictu will be discussed. Focus will be on:

-          separation techniques of complex protein mixtures using gelelectrophoreses or chromatography

-          identification techniques based on mass-spectrometry

-          discussion of different strategies for comparing protein-profiles (ICAT, iTRAQ, DIGE, O16/O18 labelling)

These theoretical lectures will be followed by practica: i) labwork (sample handling and gelelectrophoreses), ii) demonstrative practical work (2D-microcapilliary chromatography and mass-spectrometry).

Own notes, notes on Blackboard

Colignan, J.E., Dunn, B.M., Speicher, D.W., Wingfield, P.T.
Current protocols in protein science
J. Wiley & Sons, NY 2004

Xaveer Van Ostade

Departement Biomedische Wetenschappen
Campus Drie Eiken, Gebouw T, lokaal 
Universiteit Antwerpen
Universiteitsplein 1
2610 Wilrijk
 
email : xaveer.vanostade@ua.ac.be
tel: 03 265  2319

Sylvia Dewilde
Departement Biomedische Wetenschappen
Campus Drie Eiken, Gebouw T, lokaal 1.26
Universiteit Antwerpen
Universiteitsplein 1
2610 Wilrijk
 
email : sylvia.dewilde@ua.ac.be

tel: 03 265 2323

fax: 03 265 2339

Eva Geuens: eva.geuens@ua.ac.be

Lesley Tilleman : lesley.tilleman@ua.ac.be