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1.
J Mater Chem B ; 3(11): 2341-2349, 2015 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-32262064

RESUMO

Herein we report the dependency of the performance (uptake, activity, stability) of hydrophilic glucose-6-phosphate dehydrogenase (G6PDH) onto mesoporous cellular foams (MCF) grafted with aminosilanes of different chain length. The resulting hydrophobic-hydrophilic contrast was carefully evaluated by combined argon and water sorption and quantified with a newly developed hydrophilicity index suitable for the characterization of large-pore materials. The enzymatic behaviour was influenced by electrostatic and hydrophobic interactions between both reaction partners due to the creation of specific microenvironments inside the pore system. The microenvironment created by the materials with high hydrophobicity did not contribute to the beneficial electrostatic interactions of the charged amine groups by increased hydrophobic forces instead a competition of both forces was present. Thus an improved biocatalytic performance was not observed for materials with high hydrophobicity but short-chain functionalized MCFs led to highly stable and active biocatalysts.

2.
J Biol Chem ; 287(25): 21214-23, 2012 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-22544741

RESUMO

ADAM15, a disintegrin and metalloproteinase, is capable of counteracting genotoxic stress-induced apoptosis by the suppression of caspase-3 activation. A cell line expressing the membrane-bound ADAM15 without its cytoplasmic tail, however, lost this anti-apoptotic property, suggesting a crucial role of the intracellular domain as a scaffold for recruitment of survival signal-transducing kinases. Accordingly, an enhanced phosphorylation of FAK at Tyr-397, Tyr-576, and Tyr-861 was detected upon genotoxic stress by camptothecin in ADAM15-transfected T/C28a4 cells, but not in transfectants expressing an ADAM15 mutant without the cytoplasmic tail. Accordingly, a specific binding of the cytoplasmic ADAM15 domain to the C terminus of FAK could be shown by mammalian two-hybrid, pulldown, and far Western studies. In cells expressing full-length ADAM15, a concomitant activation of Src at Tyr-416 was detected upon camptothecin exposure. Cells transfected with a chimeric construct consisting of the extracellular IL-2 receptor α-chain and the cytoplasmic ADAM15 domain were IL-2-stimulated to prove that the ADAM15 tail can transduce a percepted extracellular signal to enhance FAK and Src phosphorylation. Our studies further demonstrate Src binding to FAK but not a direct Src interaction with ADAM15, suggesting FAK as a critical intracellular adaptor for ADAM15-dependent enhancement of FAK/Src activation. Moreover, the apoptosis induction elicited by specific inhibitors (PP2, FAK 14 inhibitor) of FAK/Src signaling was significantly reduced by ADAM15 expression. The newly uncovered counter-regulatory response to genotoxic stress in a chondrocytic survival pathway is potentially also relevant to apoptosis resistance in neoplastic growth.


Assuntos
Proteínas ADAM/metabolismo , Condrócitos/enzimologia , Dano ao DNA , Quinase 1 de Adesão Focal/metabolismo , Proteínas de Membrana/metabolismo , Transdução de Sinais , Proteínas ADAM/genética , Caspase 3/genética , Caspase 3/metabolismo , Linhagem Celular , Ativação Enzimática/genética , Inibidores Enzimáticos/farmacologia , Quinase 1 de Adesão Focal/genética , Humanos , Interleucina-2/genética , Interleucina-2/metabolismo , Subunidade alfa de Receptor de Interleucina-2/genética , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Proteínas de Membrana/genética , Fosforilação , Estrutura Terciária de Proteína , Quinases da Família src/antagonistas & inibidores , Quinases da Família src/genética , Quinases da Família src/metabolismo
3.
Nanoscale ; 3(3): 1234-9, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21258714

RESUMO

We have developed a new facile procedure for manufacturing crystalline thin films of SnO2 with a uniform mesoporous architecture and full crystallinity of the walls. The procedure is based on the evaporation-induced self-assembly (EISA) of prehydrolyzed tin oxide precursor directed by a commercially available Pluronic polymer. The formation of the tin oxide precursor, which can be self-assembled into a mesoporous structure, is achieved by an addition of ammonium hydroxide to a tin tetrachloride solution. The relative concentration of ammonium hydroxide as well as the duration and temperature of the hydrolysis reaction influence significantly the properties of hydrolyzed tin oxide species and the mesostructure assembled from them. The films coated from these precursor solutions and calcined at 300 °C to 400 °C exhibit a well-developed worm-like porosity with a wall to wall distance of ca. 18 nm, a surface area of up to 50 cm2 cm(-2) (corresponding to 55±5 m2 g(-1)), and high crystallinity.


Assuntos
Cristalização/métodos , Membranas Artificiais , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Compostos de Estanho/síntese química , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Tamanho da Partícula , Transição de Fase , Porosidade , Propriedades de Superfície
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