Biomedical subjects

Igor Musevic

Publications and source records attributed to Igor Musevic.

5 recordsLinked to original sources

PubMed2006

Two-dimensional nematic colloidal crystals self-assembled by topological defects.

The ability to generate regular spatial arrangements of particles is an important technological and fundamental aspect of colloidal science. We showed that colloidal particles confined to a few-micrometer-thick layer of a nematic liquid crystal form two-dimensional crystal structures that are bound by topological defects. Two basic crystalline structures were observed, depending on the ordering of the liquid crystal around the particle. Colloids inducing quadrupolar order crystallize into weakly bound two-dimensional ordered structure, where the particle interaction is mediated by the sharing of localized topological defects. Colloids inducing dipolar order are strongly bound into antiferroelectric-like two-dimensional crystallites of dipolar colloidal chains. Self-assembly by topological defects could be applied to other systems with similar symmetry.

Journal Article↗

PubMed2004

Different propensity to form amyloid fibrils by two homologous proteins-Human stefins A and B: searching for an explanation.

By using ThT fluorescence, X-ray diffraction, and atomic force microscopy (AFM), it has been shown that human stefins A and B (subfamily A of cystatins) form amyloid fibrils. Both protein fibrils show the 4.7 A and 10 A reflections characteristic for cross beta-structure. Similar height of approximately 3 nm and longitudinal repeat of 25-27 nm were observed by AFM for both protein fibrils. Fibrils with a double height of 5.6 nm were only observed with stefin A. The fibril's width for stefin A fibrils, as observed by transmission electron microscopy (TEM), was in the same range as previously reported for stefin B (Zerovnik et al., Biochem Biophys Acta 2002;1594:1-5). The conditions needed to undergo fibrillation differ, though. The amyloid fibrils start to form at pH 5 for stefin B, whereas in stefin A, preheated sample has to be acidified to pH < 2.5. In both cases, adding TFE, seeding, and alignment in a strong magnetic field accelerate the fibril growth. Visual analysis of the three-dimensional structures of monomers and domain-swapped dimers suggests that major differences in stability of both homologues stem from arrangement of specific salt bridges, which fix alpha-helix (and the alpha-loop) to beta-sheet in stefin A monomeric and dimeric forms.

Amyloid↗

PubMed2003

Structural forces near phase transitions of liquid crystals.

The structure of the fragile liquid-crystalline phases has a strong impact on the forces between bodies immersed in a liquid crystal (LC). We have equipped an atomic force microscope with a precise temperature control and measured various liquid-crystalline structural forces at temperatures close to the phase transitions. The observed forces agree well with predictions of Landau--de Gennes phenomenological theory of LCs, even at a nanoscale length. In addition to this, we have observed a molecular layer, adsorbed on the surfactant-covered glass surface, and determined its thickness and elastic properties.

Journal Article↗

PubMed2002

Human stefin B readily forms amyloid fibrils in vitro.

Human stefin B (cystatin B) is an intracellular cysteine proteinase inhibitor broadly distributed in different tissues. Here, we show that recombinant human stefin B readily forms amyloid fibrils in vitro. It dimerises and further oligomerises, starting from the native-like acid intermediate, I(N), populated at pH 5. On standing at room temperature it produces regular (over 4 microm long) fibrils over a period of several months. These have been visualised by transmission electron microscopy and atomic force microscopy. Their cross-sectional diameter is about 14 nm and blocks of 27 nm repeat longitudinally. The fibrils are smooth, of unbranched surface, consistent with findings of other amyloid fibrils. Thioflavin T fluorescence spectra as a function of time were recorded and Congo red dye binding to the fibrils was demonstrated. Adding 10% (v/v) trifluoroethanol resulted in an increased rate of fibrillation with a typical lag phase. The finding that human stefin B, in contrast to the homologue stefin A, forms amyloid fibrils rather easily should promote further studies of the protein's behaviour in vivo, and/or as a model system for fibrillogenesis.

Amyloid beta-Peptides↗

PubMed2002

Amyloid fibril formation by human stefin B in vitro: immunogold labelling and comparison to stefin A.

The mechanism by which proteins form amyloid fibrils is of high interest to the scientific community as its understanding could resolve questions relevant to conformational diseases. The structural and energetic basis of the process is still largely unknown. The main controversial issue is the co-existence of several protein conformations. Three models for the mechanism of protein fibrillogenesis have been proposed which need to be tested by experiments. In this report, amyloid fibrils grown from human stefin B (type I cystatin) are described. This physiologically relevant protein readily forms fibrils in vitro, in contrast to the homologue--human stefin A--which forms fibrils under extreme conditions only. In order to specifically label stefin B fibrils in vitro, rabbit polyclonal antibody and mouse monoclonal antibody A6/2 against human stefin B were used for immunogold labelling. Samples were examined by transmission electron microscopy. Fibrils of stefin B were strongly labelled using polyclonal antibody and Protein A gold, whereas no positive reaction was observed with monoclonal antibody A6/2.

Amyloid↗