Covalent immobilization of delipidated human serum albumin on poly(pyrrole-2-carboxylic) acid film for the impedimetric detection of perfluorooctanoic acid

The immobilization of biomolecules at screen printed electrodes for biosensing applications is still an open challenge. To enrich the toolbox of bioelectrochemists, graphite screen printed electrodes (G-SPE) were modified with an electropolymerized film of pyrrole-2-carboxilic acid (Py-2-COOH), a pyrrole derivative rich in carboxylic acid functional groups. These functionalities are suitable for the covalent immobilization of biomolecular recognition layers. The electropolymerization was first optimized to obtain stable and conductive polymeric films, comparing two different electrolytes: sodium dodecyl sulphate (SDS) and sodium perchlorate. The G-SPE modified with Py-2-COOH in 0.1 M SDS solution showed the required properties and were further tested. A proof-of-concept study for the development of an impedimetric sensor for perfluorooctanoic acid (PFOA) was carried out using the delipidated human serum albumin (hSA) as bioreceptor. The data interpretation was supported by size exclusion chromatography and small-angle X-ray scattering (SEC-SAXS) analysis of the bioreceptor-target complex and the preliminary results suggest the possibility to further develop this biosensing strategy for toxicological and analytical studies

Synthesis, structure, and characterization of 4,4′-(Anthracene-9,10-diylbis(ethyne-2,1-diyl))bis(1-methyl-1-pyridinium) Bismuth Iodide (C30H22N2)3Bi4I18, an air, water, and thermally stable 0D hybrid Perovskite with high photoluminescence ffficiency

4,4'-(Anthracene-9,10-diylbis(ethyne-2,1-diyl))bis(1-methyl-1-pyridinium) bismuth iodide (C30H22N2)3Bi4I18 (AEPyBiI) was obtained as a black powder by a very simple route by mixing an acetone solution of BiI3 and an aqueous solution of C30H22N2I2. This novel perovskite is air and water stable and displays a remarkable thermal stability up to nearly 300 °C. The highly conjugated cation C30H22N2 2+ is hydrolytically stable, being nitrogen atoms quaternarized, and this accounts for the insensitivity of the perovskite toward water and atmospheric oxygen under ambient conditions. The cation in aqueous solution is highly fluorescent under UV irradiation (emitting yellow-orange light). AEPyBiI as well is intensely luminescent, its photoluminescence emission being more than 1 order of magnitude greater than that of high-quality InP epilayers. The crystal structure of AEPyBiI was determined using synchrotron radiation single-crystal X-ray diffraction. AEPyBiI was extensively characterized using a wide range of techniques, such as X-ray powder diffraction, diffuse reflectance UV-vis spectroscopy, Fourier transform infrared (FTIR) and Raman spectroscopies, thermogravimetry-differential thermal analysis (TG-DTA), elemental analysis, electrospray ionization mass spectroscopy (ESI-MS), and photoluminescence spectroscopy. AEPyBiI displays a zero-dimensional (0D) perovskite structure in which the inorganic part is constituted by binuclear units consisting of two face-sharing BiI6 octahedra (Bi2I9 3- units). The C30H22N2 2+ cations are stacked along the a-axis direction in a complex motif. Considering its noteworthy light-emitting properties coupled with an easy synthesis and environmental stability, and its composition that does not contain toxic lead or easily oxidable Sn(II), AEPyBiI is a promising candidate for environmentally friendly light-emitting devices

The Structure of the T190M Mutant of Murine α-Dystroglycan at High Resolution: Insight into the Molecular Basis of a Primary Dystroglycanopathy

Bozzi M, Cassetta A, Covaceuszach S, et al. The Structure of the T190M Mutant of Murine α-Dystroglycan at High Resolution: Insight into the Molecular Basis of a Primary Dystroglycanopathy. PLOS ONE. 2015;10(5): e0124277

Influence of substrate on molecular order for self-assembled adlayers of CoPc and FePc

Self-assembled metal phthalocyanine thin films are receiving considerable interest due to their potential technological applications. In this study, we present a comprehensive study of CoPc and FePc thin films of about 50 nm thickness on technologically relevant substrates such as SiOx/Si, indium tin oxide (ITO) and polycrystalline gold in order to investigate the substrate induced effects on molecular stacking and crystal structure. Raman spectroscopic analysis reveals lower intensity for the vibrational bands corresponding to phthalocyanine macrocycle for the CoPc and FePc thin films grown on ITO as compared to SiOx/Si due to the higher order of phthalocyanine molecules on SiOx/Si. Atomic force microscopy analysis displays higher grain size for FePc and CoPc thin films on ITO as compared to SiOx/Si and polycrystalline gold indicating towards the influence of molecule\u2013substrate interactions on the molecular stacking. Grazing incidence X-ray diffraction reciprocal space maps reveal that FePc and CoPc molecules adopt a combination of herringbone and brickstone arrangement on SiOx/Si and polycrystalline gold substrate, which can have significant implications on the optoelectronic properties of the films due to unique molecular stacking

(3R,5S)-5(3)-Carb­oxy-3,4,5,6-tetra­hydro-2H-1,4-thia­zin-4-ium-3(5)-carboxyl­ate

The molecule of the zwitterionic title compound, C6H9NO4S, which lies on a mirror plane, shows a puckered chair conformation of the six-membered ring with the S and N atoms out of the mean plane of the other four C atoms by 0.929 (2) and 0.647 (2) Å, respectively. The ionized carboxyl group is equatorially oriented. The hydrogen-bonding network includes very short O—H⋯O [2.470 (2) Å] and N—H⋯S [3.471 (2) and 3.416 (2) Å] inter­molecular contacts

Structural flexibility of human α-dystroglycan

Dystroglycan (DG), composed of \uce\ub1 and \uce\ub2 subunits, belongs to the dystrophin-associated glycoprotein complex. \uce\ub1-DG is an extracellular matrix protein that undergoes a complex post-translational glycosylation process. The bifunctional glycosyltransferase like-acetylglucosaminyltransferase (LARGE) plays a crucial role in the maturation of \uce\ub1-DG, enabling its binding to laminin. We have already structurally analyzed the N-terminal region of murine \uce\ub1-DG (\uce\ub1-DG-Nt) and of a pathological single point mutant that may affect recognition of LARGE, although the structural features of the potential interaction between LARGE and DG remain elusive. We now report on the crystal structure of the wild-type human \uce\ub1-DG-Nt that has allowed us to assess the reliability of our murine crystallographic structure as a \uce\ub1-DG-Nt general model. Moreover, we address for the first time both structures in solution. Interestingly, small-angle X-ray scattering (SAXS) reveals the existence of two main protein conformations ensembles. The predominant species is reminiscent of the crystal structure, while the less populated one assumes a more extended fold. A comparative analysis of the human and murine \uce\ub1-DG-Nt solution structures reveals that the two proteins share a common interdomain flexibility and population distribution of the two conformers. This is confirmed by the very similar stability displayed by the two orthologs as assessed by biochemical and biophysical experiments. These results highlight the need to take into account the molecular plasticity of \uce\ub1-DG-Nt in solution, as it can play an important role in the functional interactions with other binding partners

The effect of the pathological V72I, D109N and T190M missense mutations on the molecular structure of α-dystroglycan

Dystroglycan (DG) is a highly glycosylated protein complex that links the cytoskeleton with the extracellular matrix, mediating fundamental physiological functions such as mechanical stability of tissues, matrix organization and cell polarity. A crucial role in the glycosylation of the DG α subunit is played by its own N-terminal region that is required by the glycosyltransferase LARGE. Alteration in this O-glycosylation deeply impairs the high affinity binding to other extracellular matrix proteins such as laminins. Recently, three missense mutations in the gene encoding DG, mapped in the α-DG N-terminal region, were found to be responsible for hypoglycosylated states, causing congenital diseases of different severity referred as primary dystroglycanopaties.To gain insight on the molecular basis of these disorders, we investigated the crystallographic and solution structures of these pathological point mutants, namely V72I, D109N and T190M. Small Angle X-ray Scattering analysis reveals that these mutations affect the structures in solution, altering the distribution between compact and more elongated conformations. These results, supported by biochemical and biophysical assays, point to an altered structural flexibility of the mutant α-DG N-terminal region that may have repercussions on its interaction with LARGE and/or other DG-modifying enzymes, eventually reducing their catalytic efficiency

Tin(IV) and organotin(IV) complexes containing mono or bidentate N-donor ligands. II. 4-Phenylimidazole derivatives. Crystal and molecular structure of [bis(4-phenylimidazole)trimetyltin(IV)] chloride.

A series of 2:1 adducts of the type [(L′)2RnSnX4−n]·zH2O (L′ = 4-phenylimidazole, R = Me, Et, nBu, Ph or Cy, X = I, Br or Cl, n = 0, I, 2 or 3, z = 0 or 1) has been characterized in the solid state and in solution by analyses, spectral (IR, far-IR, 1H and 13C) data and conductivity measurements. The derivatives [(L′)2(Me)3Sn]Cl (1) and [(L′)2(Me)2SnCl2] (3) react with NaClO4 in THF giving the ionic complexes [(L′)2(Me)3Sn]ClO4 and (L′)3[(Me)2Sn(ClO4)2]·(H2O)4 respectively. Whereas the triorganotin(IV) derivatives are completely dissociated in acetone solution, the diorganotin(IV) derivatives dissociate only partly and the tri- and tetrahalidetin(IV) complexes probably retain the hexacoordinate configuration. The crystal structure of [(L′)2(Me)3Sn]Cl (1) has been determined by X-ray analysis. The tin atom is coordinated to three methyl groups and two 4-phenylimidazole donors in a substantially regular trigonal bipyramidal geometry. The ionic chloride group and the two N-H moieties are involved in a hydrogen bond network

Molecular architecture of the glycogen- committed PP1/PTG holoenzyme

The delicate alternation between glycogen synthesis and degradation is governed by the interplay between key regulatory enzymes altering the activity of glycogen synthase and phosphorylase. Among these, the PP1 phosphatase promotes glycogenesis while inhibiting glycogenolysis. PP1 is, however, a master regulator of a variety of cellular processes, being conveniently directed to each of them by scaffolding subunits. PTG, Protein Targeting to Glycogen, addresses PP1 action to glycogen granules. In Lafora disease, the most aggressive pediatric epilepsy, genetic alterations leading to PTG accumulation cause the deposition of insoluble polyglucosans in neurons. Here, we report the crystallographic structure of the ternary complex PP1/PTG/carbohydrate. We further refine the mechanism of the PTG-mediated PP1 recruitment to glycogen by identifying i) an unusual combination of recruitment sites, ii) their contributions to the overall binding affinity, and iii) the conformational heterogeneity of this complex by in solution SAXS analyses