MPŠ MP&Scaron MP&Scaron MP&Scaron Avtorji

Jožef Stefan
Postgraduate School

Jamova 39
SI-1000 Ljubljana

Phone: +386 1 477 31 00
Fax: +386 1 477 31 10


Course Description

Protein and Cell Engineering


Nanosciences and Nanotechnologies, third-level study programme


prof. dr. Jože Pungerčar


A student is expected to acquire in-depth knowledge and understanding of structure-function relationships in proteins. He/she learns how to independently design appropriate experiments and to use different approaches in protein engineering. The student also learns how to exploit various cell systems for studying and understanding the biological roles of proteins of interest.


• Protein engineering: historical view, basic concepts and terms, rational design and directed evolution
• Mutagenesis: various approaches and common techniques
• Unlimited potential of cell engineering
• Use of various protein tags and markers
• Design of site-directed mutations on a selected protein molecule
• Introduction of mutation(s) at the nucleotide level
• Production of a mutant recombinant protein and its basic characterization
• Preparation of a selected labeled protein in mammalian cells
• Analysis of the obtained results
• Preparation of a final seminar work

Course literature:

• Ausubel, F.M. et al. /Eds./ (2007): Current Protocols in Molecular Biology. John Wiley and Sons, Inc., New York, USA; Continuously updated.

• Robertson, D.E. and Noel, J.P. /Eds./ (2004): Protein Engineering. In: Methods in Enzymology, Vol. 388, ElsevierAcademic Press, Amsterdam, The Netherlands.

• Al-Rubeai, M. /Ed./ (1999): Cell Engineering, Vol. 1. Springer Verlag, New York, USA.

• Al-Rubeai, M. and Fussenegger, M. /Eds./ (2007): Cell Engineering, Vol. 5 (Systems Biology), Springer Verlag, New York, USA.

• Current research and review articles from the field.

Significant publications and references:

• Jenko Pražnikar, Z., Kovačič, L., Rowan, E.G., Romih, R., Rusmini, P., Poletti, A., Križaj, I. and Pungerčar, J. (2008): A presynaptically toxic secreted phospholipase A2 is internalized into motoneuron-like cells where it is rapidly translocated into the cytosol. Biochim. Biophys. Acta – Mol. Cell Res. 1783, 1129–1139.

• Petan, T., Križaj, I. and Pungerčar, J. (2007): Restoration of enzymatic activity in a Ser-49 phospholipase A2 homologue decreases its Ca(2+)-independent membrane damaging activity and increases its toxicity. Biochemistry 46, 12795–12809.

• Petan, T., Križaj, I., Gelb, M.H. and Pungerčar, J. (2005): Ammodytoxins, potent presynaptic neurotoxins, are also highly efficient phospholipase A2 enzymes. Biochemistry 44, 12535–12545.

• Prijatelj, P., Šribar, J., Ivanovski, G., Križaj, I., Gubenšek, F. and Pungerčar, J. (2003): Identification of a novel binding site for calmodulin in ammodytoxin A, a neurotoxic group IIA phospholipase A2. Eur. J. Biochem. 270, 3018–3025.

• Petan, T., Križaj, I., Gubenšek, F. and Pungerčar, J. (2002): Phenylalanine-24 in the N-terminal region of ammodytoxins is important for both enzymic activity and presynaptic toxicity. Biochem. J. 363, 353–358.

• Francky, A., Mozetič Francky, B., Štrukelj, B., Gruden, K., Ritonja, A., Križaj, I., Kregar, I., Pain, R.H. and Pungerčar, J. (2001): A basic residue at position 36p of the propeptide is not essential for the correct folding and subsequent autocatalytic activation of prochymosin. Eur. J. Biochem. 268, 2362–2368.

• Anderluh, G., Barlič, A., Podlesek, Z., Maček, P., Pungerčar, J., Gubenšek, F., Zecchini, M.L., Dalla Serra, M. and Menestrina, G. (1999): Cysteine-scanning mutagenesis of an eukaryotic pore-forming toxin from sea anemone. Topology in lipid membranes. Eur. J. Biochem. 263, 128–136.


• Seminar work and oral exam

Students obligations:

• Practical laboratory work
• Seminar and oral examination