A5079931608- Protease and Inhibitor Mechanisms
- Peptidase Inhibition and Analysis
- Bone Metabolism and Diseases
- Biochemical and Structural Characterization
- Cell Adhesion Molecules Research
- Venomous Animal Envenomation and Studies
- Cellular transport and secretion
- Natural product bioactivities and synthesis
- Signaling Pathways in Disease
- Insect Resistance and Genetics
- Calpain Protease Function and Regulation
- X-ray Diffraction in Crystallography
- Crystallization and Solubility Studies
- Crystallography and molecular interactions
- SARS-CoV-2 and COVID-19 Research
- Bone and Dental Protein Studies
- Studies on Chitinases and Chitosanases
- Biochemical and Molecular Research
- Transgenic Plants and Applications
- Neuropeptides and Animal Physiology
- HIV/AIDS drug development and treatment
- Chemical Synthesis and Analysis
- Lipid Membrane Structure and Behavior
- Trace Elements in Health
- Synthesis and Characterization of Pyrroles
University of Ljubljana
2015-2025
University of Zurich
2012-2014
Jožef Stefan Institute
2005-2010
Proteolytic degradation of elastic fibers is associated with a broad spectrum pathological conditions such as atherosclerosis and pulmonary emphysema. We have studied the interaction between elastins human cysteine cathepsins K, L, S, which are known to participate in elastinolytic activity vivo. The enzymes showed distinctive preferences degrading from bovine neck ligament, aorta, lung. Different susceptibility these proteolysis was attributed morphological differences observed by scanning...
Zinc pyrithione (1a), together with its analogues 1b-h and ruthenium complex 2a, were synthesised evaluated for the stability in biologically relevant media anti-SARS-CoV-2 activity. revealed potent vitro inhibition of cathepsin L (IC50=1.88 ± 0.49 µM) PLPro (IC50=0.50 0.07 µM), enzymes involved SARS-CoV-2 entry replication, respectively, as well antiviral replication properties an ex vivo system derived from primary human lung tissue. complexes expressed comparable inhibition. On contrary,...
The human cysteine peptidase cathepsin K is a key enzyme in bone homoeostasis and other physiological functions. In the present study we investigate mechanism of action at plasma pH its regulation by modifiers that bind outside active site. We show fluctuates between multiple conformations are differently susceptible to macromolecular inhibitors can be manipulated varying ionic strength medium. behaviour vitro described presence two discrete with distinctive kinetic properties different...
Summary Caspases are a family of cysteine‐dependent proteases known to be involved in the process programmed cell death metazoans. Recently, cyanobacteria were also found contain caspase‐like proteins, but their existence has only been identified silico up now. Here, we present first experimental characterisation prokaryotic caspase homologue. We have expressed putative gene MaOC 1 from toxic bloom‐forming cyanobacterium M icrocystis aeruginosa PCC 7806 E scherichia coli . Kinetic showed...
Abstract From the point of view medicinal chemistry, thiosemicarbazones are interesting molecules as they exhibit various biological activities, such antitumor, antifungal, antibacterial, antiviral, and antileukemic properties, their synthesis is deeply investigated. However, using 2‐methylthiosemicarbazide a starting compound to prepare new 1,2,4‐triazolidine‐3‐thione compounds via 2‐methylthiosemicarbazone cyclization has not been reported yet. Therefore, fifteen were synthesized during...
Multidomain proteins are gaining increasing consideration for their puzzling, flexible utilization in nature. The presence of the characteristic thyroglobulin type-1 (Tg1) domain as a protein module variety multicellular organisms suggests pivotal roles this building block. To gain insight into evolution Tg1 domains, we performed searches protein, expressed sequence tag, and genome databases. domains were found to be Metazoa specific, retrieved total 170 domain–containing sequences. Their...
Secreted modular calcium-binding proteins 1 and 2 (SMOC-1 SMOC-1) are extracellular calcium- binding belonging to the BM-40 family of proteins. In this work we have identified a highly basic region in (EC) domain SMOC-1 similar other known glycosaminoglycan-binding motifs. Size-exclusion chromatography shows that full length as well its C-terminal EC alone bind heparin heparan sulfate, but not related chondroitin sulfate or dermatan glycosaminoglycans. Intrinsic tryptophan fluorescence...
The cysteine protease CP14 has been identified as a central component of molecular module regulating programmed cell death in plant embryos. belongs to distinct subfamily papain-like proteinases which no representative characterized thoroughly date. However, it proposed that is cathepsin H-like protease. We have now produced recombinant Nicotiana benthamiana (NbCP14) lacking the C-terminal granulin domain. As typical for proteinases, NbCP14 undergoes rapid autocatalytic activation when...
The tobacco-related plant Nicotiana benthamiana is gaining interest as a versatile host for the production of monoclonal antibodies and other protein therapeutics. However, susceptibility plant-derived recombinant proteins to endogenous proteolytic enzymes limits their use biopharmaceuticals. We have now identified two previously uncharacterized N. proteases with high antibody-degrading activity, papain-like cysteine proteinases NbCysP6 NbCysP7. Both are capable hydrolysing wide range...
Abstract Toxic cyanobacterial blooms are a rich source of metabolites having variety biological activities. Two main groups cyclic peptides, depsipeptides and ureido linkage-containing reportedly inhibit serine peptidases. We characterised their inhibitory properties against selected peptidases investigated influence on cell viability. The depsipeptide planktopeptin BL1125 is strong linear competitive tight-binding inhibitor leukocyte ( K i =2.9 n m ) pancreatic =7.2 elastase also...
Targeting allosteric sites is gaining increasing recognition as a strategy for modulating the activity of enzymes, especially in drug design. Here we investigate mechanisms regulation cathepsin K representative cysteine cathepsins and promising target treatment osteoporosis. Eight novel modifiers are identified by computational targeting predicted on surface enzyme. All act via hyperbolic kinetic presence low molecular mass substrates, expected effectors. Two compounds have sizable effects...
The tobacco-related plant species Nicotiana benthamiana has recently emerged as a versatile expression platform for the rapid generation of recombinant biopharmaceuticals, but product yield and quality frequently suffer from unintended proteolysis. Previous studies have highlighted that protein fragmentation in plants involves papain-like cysteine proteinases (PLCPs). For this reason, we now characterized two major N. PLCPs detail: aleurain-like protease (NbALP) cathepsin B (NbCathB). As...
Abstract The genome of the model plant Arabidopsis thaliana encodes three paralogues papain-like cysteine proteinase cathepsin B (AtCathB1, AtCathB2 and AtCathB3), whose individual functions are still largely unknown. Here we show that a mutated splice site causes severe truncations AtCathB1 polypeptide, rendering it catalytically incompetent. By contrast, AtCathB3 effective proteases which display comparable hydrolytic properties share most their substrate specificities. Site-directed...
Several members of the papain-like peptidase family have ability to degrade collagen molecules by cleaving within triple helix region this difficult substrate. A common denominator these peptidases is their cleave substrates with Pro in P2 position. In humans, cathepsin K best-known collagenase. Here, we investigate collagenolytic activity human L, which closely related K. We show that, despite lacking proline specificity, L efficiently cleaves type I and produces a cleavage pattern similar...