Interfering with the inhibitory mechanism of serpins: crystal structure of a complex formed between cleaved plasminogen activator inhibitor type 1 and a reactive-centre loop peptide

BACKGROUND: Plasminogen activator inhibitor type 1 (PAI-1) is an important endogenous regulator of the fibrinolytic system. Reduction of PAI-1 activity has been shown to enhance dissolution of blood clots. Like other serpins, PAI-1 binds covalently to a target serine protease, thereby irreversibly inactivating the enzyme. During this process the exposed reactive-centre loop of PAI-1 is believed to undergo a conformational change becoming inserted into beta sheet A of the serpin. Incubation with peptides from the reactive-centre loop transform serpins into a substrate for their target protease. It has been hypothesised that these peptides bind to beta sheet A, thereby hindering the conformational rearrangement leading to loop insertion and formation of the stable serpin-protease complex. RESULTS: We report here the 1.95 A X-ray crystal structure of a complex of a glycosylated mutant of PAI-1, PAI-1-ala335Glu, with two molecules of the inhibitory reactive-centre loop peptide N-Ac-TVASS-NH2. Both bound peptide molecules are located between beta strands 3A and 5A of the serpin. The binding kinetics of the peptide inhibitor to immobilised PAI-1-Ala335Glu, as monitored by surface plasmon resonance, is consistent with there being two different binding sites. CONCLUSIONS: This is the first reported crystal structure of a complex formed between a serpin and a serpin inhibitor. The localisation of the inhibitory peptide in the complex strongly supports the theory that molecules binding in the space between beta strands 3A and 5A of a serpin are able to prevent insertion of the reactive-centre loop into beta sheet A, thereby abolishing the ability of the serpin to irreversibly inactivate its target enzyme. The characterisation of the two binding sites for the peptide inhibitor provides a solid foundation for computer-aided design of novel, low molecular weight PAI-1 inhibitors.     

Mechanical properties of wollastonite‐reinforced polypropylene composites modified with SEBS and SEBS‐g‐MA elastomers

Mechanical properties of isotactic polypropylene/wollastonite/styrene rubber block copolymers (iPP/wollastonite/SRBC) composites were studied as a function of elastomeric poly(styrene‐b‐ethylene‐co‐butylene‐b‐styrene) triblock copolymer (SEBS) and SEBS grafted with maleic anhydride (SEBS‐g‐MA) content from 0 to 20 vol%. Microphase morphology was stronger influenced by SRBC elastomers than by different wollastonite types. Higher encapsulation ability of SEBS‐g‐MA than SEBS caused more expressive core‐shell morphology and consequently higher notched impact strength as well as yield parameters, but lower Young's modulus. Higher ductility of the composites with SEBS than with SEBS‐g‐MA has been primarily caused by better miscibility of the polypropylene chains with SEBS molecules. Surface properties of components and adhesion parameters also indicated that adhesion at SEBS‐g‐MA/wollastonite interface, which was stronger than the one at the SEBS/wollastonite interface, influenced higher encapsulation of wollastonite particles by SEBS‐g‐MA. POLYM. ENG. SCI., 47:1873–1880, 2007. © 2007 Society of Plastics Engineers     

Morphology and mechanical properties of poly(propylene)/polystyrene blends compatibilized with polystyrene-block-poly(ethylene-co-propylene)

Compatibilizing effects of diblock copolymer polystyrene-block-poly(ethylene-co-propylene) (SEP) on the morphology and mechanical properties of immiscible blends of poly(propylene) (PP) and polystyrene (PS) were investigated. Notched impact strength, yield stress, elongation at yield and Young's modulus were determined as a function of different weight ratios of PP and PS and different amounts of added SEP as well. Scanning electron microscopy revealed a two-phase morphology of PP/PS blends, which exhibit poor mechanical properties. Even 2,5 wt.-% of SEP added to PP/PS blends can improve the notched impact strength and elongation at yield compared to non-compatibilized PP/PS blends. 10 wt.-% of SEP compatibilizer converted the brittle PP/PS blend to quite impactresistant polymeric material. Mechanical properties were improved because of the morphological changes and increased interfacial adhesion as a result of SEP localization between PP and PS phases. An analysis of yield stress data in terms of theoretical models showed that yield stress values of binary PP/PS blends can be predicted with Nielsen's model.     

Composition and seasonal variation of fatty acids of <Emphasis Type="Italic">Diplodus vulgaris</Emphasis> L. from the Adriatic Sea

Muscle tissue from the common two-banded sea bream Diplodus vulgaris L. originating from the Adriatic Sea, Croatia, was analyzed. The FA composition of neutral (TAG) and polar (PE, PC, PI/PS) lipid classes was determined, as well as the lipid and water contents during winter and summer periods. Both the total lipid and water contents were higher in the winter period. We identified 16 different FA. The major constituents of the total FA in both seasons were saturates: palmitic (16∶0) and stearic acids (18∶0); monoenes: oleic (18∶1n−9) and palmitoleic acids (16∶1n−7); and polyunsaturates: arachidonic acid (20∶4n−6), EPA (20∶5n−3), and DHA (22∶6n−3), but their amounts and ratios differed significantly between the two seasons and between lipid fractions. The FA composition showed a noticeable pattern of seasonality that reflected fluctuations mainly in TAG. The diminution of the monounsaturated FA content in the summer was clearly followed by an increase in PUFA content. Diplodus vulgaris is a good source of natural n−3 PUFA and would therefore be suitable for inclusion in highly unsaturated low-fat diets.     

Fatigue crack propagation and damage evolution in PBT-GF and SAN-GF

Our earlier investigations of fatigue behavior in PBT-GF and SAN-GF with different fiber lengths have shown that fatigue crack propagation (FCP) can be described in terms of elastic-plastic fracture mechanics. In this work it is shown that the influence of structural material parameters on the resistance to FCP correlates with the extent of energy dissipation at the crack tip. With increasing fiber length, the zone of energy dissipation is increased. By means of microscopic investigations, the prevailing damage in the zone of energy dissipation is identified as micro cracks in the matrix.     

Grinding induced effects in plasma sprayed zirconia coatings

The effect of grinding on the surface layer properties of ceria and yttria partially stabilized zirconia plasma-sprayed coatings (CePSZ, YPSZ, respectively) has been studied by X-ray diffraction methods. For this purpose, the modified model of line broadening analysis has been derived. The model considers elastic anisotropic properties along with more random paracrystal imperfections, both affecting X-ray line broadening. Grinding-induced microstructural changes were also studied using an estimation from the quantitative Orientation Distribution Function (ODF) texture. It was concluded, based on this work, that CePSZ ceramic is less mechanically stable compared to YPSZ. Consequently, more beneficial mechanical properties of a ground surface layer can be expected for CePSZ plasma-sprayed coatings.     

Hydrophobicity of Thin Films of Compounds of Low‐Electronegativity Metals

Hydrophobic materials capable of withstanding harsh conditions are required for various applications. Here, we show that oxides and nitrides of various low‐electronegativity metals are hydrophobic hard ceramics. We attribute their hydrophobicity to low Lewis acidity of the low‐electronegativity cations implying a low ability of the cations on the surface to form coordinate bonds with water oxygen anions. Furthermore, we observe a systematically stronger hydrophobic behavior of nitrides compared with the corresponding oxides, which we attribute to nitrogen being a poorer Lewis base than oxygen due to a reduced number of lone pairs of electrons, implying a lower ability of nitrogen anions on the surface to form hydrogen bonds with water hydrogen cations. Most of the oxides and nitrides investigated exhibit high values of hardness. Therefore, oxides and nitrides of low‐electronegativity metals should find application as hydrophobic materials in harsh conditions.     

Sulphidation and oxidation of the Ni22Cr10Al1Y alloy in H2/H2S and SO2 atmospheres at high temperatures

The Ni22Cr10Al1Y alloy was exposed in H₂/H₂S gas mixture under the sulphur pressure 10^–3 and 1 Pa as well as in SO₂ at 1173 and 1273 K. At p_s = 1 Pa the sulphidation rate was relatively high and the reaction obeyed the linear rate law. Under these conditions a nickel/nickel sulphide eutectic was formed. At p_s = 10^–3 Pa nickel sulphides became unstable and the sulphidation rate was significantly lower. The reaction obeyed the parabolic rate law. The oxidation rate of the alloy in SO₂ was lower than that in any of the H₂/H₂S atmospheres. The sulphide scales formed during sulphidation in H₂/H₂S had complex microstructures and compositions, with sulphospinel and sulphide phases being present, e.g. NiCr₂S₄, Ni₃S₂, Cr_xS_y. As the temperature increased and the sulphur pressure decreased, these phases were replaced by the chromium-rich sulphide phase. Various oxides formed during oxidation of the alloy in SO₂.     

Polypropylene/silica micro‐ and nanocomposites modified with poly(styrene‐b‐ethylene‐co‐butylene‐b‐styrene)

The effects of different silica loadings and elastomeric content on interfacial properties, morphology and mechanical properties of polypropylene/silica 96/4 composites modified with 5, 10, 15, and 20 vol % of poly(styrene‐b‐ethylene‐co‐butylene‐b‐styrene) SEBS added to total composite volume were investigated. Four silica fillers differing in size (nano‐ vs. micro‐) and in surface properties (untreated vs. treated) were chosen as fillers. Elastomer SEBS was added as impact modifier and compatibilizer at the same time. The morphology of ternary polymer composites revealed by light and scanning electron microscopies was compared with morphology predicted models based on interfacial properties. The results indicated that general morphology of composite systems was determined primarily by interfacial properties, whereas the spherulitic morphology of polypropylene matrix was a result of two competitive effects: nucleation effect of filler and solidification effect of elastomer. Tensile and impact strength properties were mainly influenced by combined competetive effects of stiff filler and tough SEBS elastomer. Spherulitic morphology of polypropylene matrix might affect some mechanical properties additionally. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41486.