He is interested in protein-membrane interactions. He is studying how proteins interact with lipid membranes at the molecular level. Specifically, he studies pore forming proteins that are able to perforate lipid membranes in a multi-step process involving membrane binding, oligomerization and pore formation. He studies important regulators of immune system (perforin), bacterial virulence factors (listeriolysin), Nep1-like proteins and proteins similar to toxins from sea anemones (actinoporin-like proteins). He employs various biochemical and biophysical approaches with an emphasis on surface plasmon resonance, which allows studying molecular interactions. He is developing and improving protocols for the study of protein binding to lipid membranes.
1994: B.Sc. in Biology: Molecular cloning and partial gene analysis of long-nosed viper (Vipera ammodytes ammodytes L.) phospholipases A2. (supervisor: prof. dr. Igor Kregar)
1998: PhD in Biological Sciences: Protein engineering and mechanism of action of equinatoxin II from the sea anemone Actinia equina L. (supervisor: prof. dr. Peter Maček, co-supervisor prof. dr. Jože Pungerčar)
Research Subjects: Membranes and transport / Proteins / Biochemistry / Structural biology / Biophysics
Research Interests: Natural toxins / Molecular interactions / Protein-membrane interactions / Surface plasmon resonance
The complex structure, great stability, and diversity of natural pore-forming proteins (PFPs) have inspired their use in various biotechnological applications. A label-free single-molecule nanopore sensing technique is based on the detection of electric current flowing through a biological nanopore inserted into an electrically non-conductive lipid or artificial membrane. When electroosmotic and electrophoretic forces favor the translocation of the analyte molecule through the nanopore, this leads to the occurrence of electrical current blockades. In the optimized system, the blockades are analyte-specific, which allows their discrimination in more complex samples. Protein detection (protein fingerprinting) and amino acid sequence reads are gaining attention now that nanopore DNA sequencing has become a powerful third-generation sequencing method in many laboratories. Recent results highlight peptides translocation, single amino acid differentiation, post-translational modification detection, and assisted protein translocation, which makes nanopore-based protein detection a promising alternative to conventional mass spectrometry. Efficient detection of proteins at single amino acid resolution requires new nanopores with novel dimensions and biochemical properties. Nanopores that stably insert into artificial membranes and enable high-throughput detection of medically relevant proteins are particularly attractive. Our research focuses on pore-forming toxins (PFTs), in particular actinoporins and their homologs, Nep1-like proteins, and larger cholesterol-dependent cytolysins. In addition to traditional genetic approaches, we are also investigating the incorporation of unnatural amino acids into the pore sensing region, which may broaden the potential spectrum of analysis.