Enzymes Immobilized In Langmuir-Blodgett films: Why determining the surface properties in Langmuir monolayer is important?

In this review we discuss about the immobilization of enzymes in Langmuir-Blodgett films in order to determine the catalytic properties of these biomacromolecules when adsorbed on solid supports. Usually, the conformation of enzymes depends on the environmental conditions imposed to them, including the chemical composition of the matrix, and the morphology and thickness of the film. In this review, we show an outline of manuscripts that report the immobilization of enzymes as LB films since the 1980's, and also some examples of how the surface properties of the floating monolayer prepared previously to the transfer to the solid support are important to determine the efficiency of the resulting device.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Univ Fed Sao Paulo, Inst Environm Chem & Pharmaceut Sci, Rua Sao Nicolau 210, BR-09913030 Diadema, SP, BrazilUniv Fed Sao Paulo, Inst Environm Chem & Pharmaceut Sci, Rua Sao Nicolau 210, BR-09913030 Diadema, SP, BrazilFAPESP: 2015/10851-0Web of Scienc

Probing the interaction between heparan sulfate proteoglycan with biologically relevant molecules in mimetic models for cell membranes: A Langmuir film study

Investigating the role of proteoglycans associated to cell membranes is fundamental to comprehend biochemical process that occurs at the level of membrane surfaces. in this paper, we exploit syndecan-4, a heparan sulfate proteoglycan obtained from cell cultures, in lipid Langmuir monolayers at the air-water interface. the monolayer served as a model for half a membrane, and the molecular interactions involved could be evaluated with tensiometry and vibrational spectroscopy techniques. Polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS) employed in a constant surface pressure regime showed that the main chemical groups for syndecan-4 were present at the air-water interface. Subsequent monolayer decompression and compression showed surface pressure-area isotherms with a large expansion for the lipid monolayers interacting with the cell culture reported to over-express syndecan-4, which was also an indication that the proteoglycan was inserted in the lipid monolayer. the introduction of biological molecules with affinity for syndecam-4, such as growth factors, which present a key role in biochemical process of cell signaling, changed the surface properties of the hybrid film, leading to a model, by which the growth factor binds to the sulfate groups present in the heparan sulfate chains. the polypeptide moiety of syndecan-4 responds to this interaction changing its conformation, which leads to lipid film relaxation and further monolayer condensation. (C) 2012 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, Diadema, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, São Paulo, SP, BrazilUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, Diadema, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, São Paulo, SP, BrazilWeb of Scienc

The lipid composition of a cell membrane modulates the interaction of an antiparasitic peptide at the air-water interface

The antiparasitic property of peptides is believed to be associated with their interactions with the protozoan membrane, which calls for research on the identification of membrane sites capable of peptide binding. in this study we investigated the interaction of a lipophilicglutathioine peptide known to be effective against the African Sleeping Sickness (ASS - African Trypanosomiasis) and cell membrane models represented by Langmuir monolayers. It is shown that even small amounts of the peptide affect the monolayers of some phospholipids and other lipids, which points to a significant interaction. the latter did not depend on the electrical charge of the monolayer-forming molecules but the peptide action was particularly distinctive for cholesterol + sphingomyelin monolayers that roughly resemble rafts on a cell membrane. Using in situ polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), we found that the orientation of the peptide is affected by the phospholipids and dioctadecyldimethylammonium bromide (DODAB), but not in monolayers comprising cholesterol + sphingomyelin. in this mixed monolayer resembling rafts, the peptide still interacts and has some induced order, probably because the peptide molecules are fitted together into a compact monolayer. Therefore, the lipid composition of the monolayer modulates the interaction with the lipophilic glutathioine peptide, and this may have important implications in understanding how the peptide acts on specific sites of the protozoan membrane. (C) 2011 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)rede nBioNet (Brazil)Univ Estadual Paulista, Fac Ciencias & Letras Assis, Assis, SP, BrazilUniv São Paulo, Inst Fis Sao Carlos, Sao Carlos, SP, BrazilUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, Diadema, SP, BrazilManchester Metropolitan Univ, Sch Biol Chem & Hlth Sci, Manchester M15 6BH, Lancs, EnglandUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, Diadema, SP, BrazilWeb of Scienc

Surface chemistry and spectroscopy studies on 1,4-naphthoquinone in cell membrane models using Langmuir monolayers

Investigating the role of drugs whose pharmaceutical activity is associated with cell membranes is fundamental to comprehending the biochemical processes that occur on membrane surfaces. in this work, we examined the action of 1,4-naphthoquinone in lipid Langmuir monolayers at the air-water interface, which served as a model for half of a membrane, and investigated the molecular interactions involved with tensiometry and vibrational spectroscopy. the surface pressure-area isotherms exhibited a noticeable shift to a lower area in relation to 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dihexadecanoyl-sn-glycero-3-phospho-L-serine (DPPS) lipid monolayers, which indicated a disruption of the monolayer structure and solubilisation of the lipids towards the aqueous subphase. To better correlate to the action of this drug in biological membrane events, cell cultures that represented tumorigenic and non-tumorigenic cells were spread onto the air-water interface, and 1,4-naphthoquinone was then incorporated. While only slight changes were observed in the non-tumorigenic cells upon drug incorporation, significant changes were observed in the tumorigenic cells, on which the organisation of the Langmuir monolayers was disrupted as evidenced by tensiometry and vibrational spectroscopy. This work then shows that this drug interacts preferentially for specific surfaces. in simplified models, it has a higher effect for the negative charged DPPS rather than the zwitterionic DPPC; and for complex cell cultures, 1,4-naphthoquinone presents a more significant effect for that representing tumorigenic cells. (c) 2013 Elsevier Inc. All rights reserved.Universidade Federal de São Paulo, Diadema, SP, BrazilUniversidade Federal de São Paulo, Diadema, SP, BrazilWeb of Scienc

Chitosan in nanostructured thin films

This review paper brings an overview of the use of chitosans in nanostructured films produced with the Langmuir-Blodgett (LB) or the electrostatic layer-by-layer (LbL) techniques, with emphasis on their possible applications. From a survey in the literature one may identify three main types of study with chitosan in nanostructured films. First, the interaction between chitosans and phospholipid Langmuir monolayers has been\ud investigated for probing the mechanisms of chitosan action in their biological applications, with the monolayers serving as cell membrane models. In the second type, chitosan serves as a matrix for immobilization of biomolecules in LB as well as in LbL films, for which chitosan is suitable to help preserve the bioactivity of such biomolecules\ud for long periods of time even in dry, solid films. An important application of these chitosan-containing films is in sensing and biosensing. The third type of study involves exploiting the mechanical and biocompatibility properties of chitosan in producing films with enhanced properties, for example, for tissue engineering. It is emphasized\ud that chitosans have been proven excellent building blocks to produce films with controlled molecular architecture, allowing for synergy between distinct materials. We also discuss the prospects of the field, following a critical review of the latest developments in nanostructured chitosan films.FAPESPCNPqCAPE

Innovative low temperature plasma approach for deposition of alumina films

Alumina films were deposited from a new plasma method using aluminum acetylacetonate (AAA) powder as precursor. The AAA was sputtered in argon and oxygen plasma mixtures. It was investigated the effect of the oxygen proportion (O2%) on the properties of the coatings. Deposition rate was derived from the layer height measured by profilometry. The elemental composition and molecular structure of the films were determined by Rutherford backscattering and infrared spectroscopies, respectively. Grazing incidence X-ray diffraction was used to investigate the microstructure of the films while hardness was determined by nanoindentation technique. Inspections on the surface morphology and on the film composition were conducted associating scanning electron microscopy and energy dispersive spectroscopy. Incorporation of oxygen affects the plasma kinetics and consequently the properties of the coatings. As moderated concentrations of oxygen ( 25%) are incorporated, the structure become rich in metallic aluminum with carbon rising at low proportions. The deposited layer is not homogeneous in thickness once the chemical composition of the precursor is changed by the action of the reactive oxygen plasma. Oxygen ablation on the film surface also contributes to the lack of homogeneity of the structure, especially as high oxygen proportions are imposed. Hardness data (0.5-2.0 GPa) corroborated the idea of an amorphous structure. Based on the results presented here it was possible to identify the oxygen concentration in the plasma atmosphere which mostly removed organics while preserving the stoichiometric alumina precipitation, subject of great relevance as one considers the reduction in the energy necessary for the creation of fully oxide coatings.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Estadual Paulista Laboratório de Plasmas TecnológicosUniversidade Federal de São Paulo (UNIFESP) Departamento de Ciências Exatas e da TerraUniversidade de São Paulo Departamento de Física NuclearUniversidade de São Paulo Departamento de Física AplicadaUNIFESP, Depto. de Ciências Exatas e da TerraSciEL

Molecular-level interactions of an azopolymer and poly(dodecylmethacrylate) in mixed Langmuir and Langmuir-Blodgett films for optical storage

The applicability of azopolymers in optical storage can be extended through the use of nanostructured films produced with the Langmuir–Blodgett (LB) technique, but the film properties need to be optimized since these polymers generally do not form stable Langmuir films to be transferred onto solid substrates. Here, photoinduced birefringence was investigated for mixed Langmuir–Blodgett films from the homopolymers 4-[N-ethyl-N-(2-methacryloxyethyl)]-4′-nitroazobenzene (HPDR1-MA) and poly(dodecylmethacrylate) (HPDod-MA). The interactions between these polymers were studied in Langmuir and LB films. Surface pressure–area isotherms pointed to molecular-level interactions for proportions of 51 mf%, 41 mf% and 31 mf% of HPDR1-MA. Phase segregation was not apparent in the BAM images, in which the morphology of the blend film was clearly different from that of the Langmuir films of neat homopolymers. Through PM-IRRAS, we noted that the interaction between the azopolymer and HPDod-MA affected the orientation of carbonyl groups. Strong interactions for the mixture with 41 mf% of poly(dodecylmethacrylate) led to stable Langmuir films that were transferred onto solid supports as LB films. The photoinduced birefringence of 101-layer mixed LB films show features that make these films useful for optical storage, with the advantage of short writing times in comparison to other azopolymer films.CAPESFAPESPCNP

Enzymes immobilized in Langmuir-Blodgett films: Why determining the surface properties in Langmuir monolayer is important?

ABSTRACT In this review we discuss about the immobilization of enzymes in Langmuir-Blodgett films in order to determine the catalytic properties of these biomacromolecules when adsorbed on solid supports. Usually, the conformation of enzymes depends on the environmental conditions imposed to them, including the chemical composition of the matrix, and the morphology and thickness of the film. In this review, we show an outline of manuscripts that report the immobilization of enzymes as LB films since the 1980’s, and also some examples of how the surface properties of the floating monolayer prepared previously to the transfer to the solid support are important to determine the efficiency of the resulting device

Influence of the glycosylphosphatidylinositol anchor in the immobilization of rat osseous plate alkaline phosphatase immobilized in biomimetic systems: phospholipid Langmuir monolayers and Langmuir-Blogett films

Nesse trabalho, estudou-se a incorporação da fosfatase alcalina de placa óssea de ratos em dois tipos de sistemas modelo que mimetizassem uma membrana celular: as monocamadas de Langmuir e os filmes Langmuir-Blodgett (LB), ambos formados com fosfolipídios. No intuito de se investigar o papel da âncora de glicosilfosfatidilinositol (GFI), covalentemente ligada ao grupo carboxi-terminal da cadeia polipeptídica, duas formas da enzima foram estudadas: uma com a âncora GFI intacta, solubilizada por ação de um tensoativo não-iônico, e a outra sem a parte hidrofóbica dessa âncora, clivada por ação de uma fosfolipase específica. A primeira forma enzimática foi chamada de FAT (fosfatase alcalina solubilizada por tensoativo), e a segunda de FAC (fosfatase alcalina clivada). Diferenças marcantes na atividade superficial foram observadas entre as duas formas. Comparativamente, a forma FAT adsorve mais rapidamente à interface ar/água que a forma FAC, que mostra um tempo de indução significativo. A incorporação da forma FAT às monocamadas de ácido dimiristoilfosfatídico (DMPA) também ocorre mais rapidamente que a forma FAC. No entanto, o uso de alta força iônica acelerou a adsorção da forma FAC à interface ar/água (com ou sem DMPA). Uma pressão de superfície de exclusão de 20mN/m foi encontrado para a forma FAC, enquanto para a forma FAT, essa pressão representa uma mudança no perfil das curvas pressão x tempo. Isso revelou que, devido à presença da âncora GFI, essa forma enzimática é capaz de incorporar às monocamadas de DMPA, mesmo em altas pressões de superfície. Isotermas superficiais de monocamadas mistas de DMPA e fosfatase alcalina também mostraram diferentes perfis para duas formas enzimáticas estudadas. Enquanto a forma FAT provoca uma alteração na compressibilidade em pressões de até 20mN/m, a forma FAC desloca a curva pressão x área para áreas mais elevadas. Tal fato foi explicado pelo fato da forma FAT incorporar na monocamada preferencialmente com a âncora GFI posicionada entre as cadeias apolares do DMPA, enquanto a forma FAC deva incorporar a cadeia polipeptídica à interface lipídica . Microscopias de fluorescência e no ângulo de Brewster revelaram que a forma FAT provoca agregação espontânea do DMPA na interface ar/água, levando a uma microeterogeneidade, na qual três fases distintas podem ser observadas. A obtenção de espectros de infravermelho na interface ar/água, associada com medidas de atividade catalítica in situ, revelou que a atividade da fosfatase alcalina é modulada pela capacidade de empacotamento interfacial, medida pelo módulo de compressibilidade superficial. Em 20mN/m, há uma reorganização molecular na interface, o que vai restringir a flexibilização da cadeia polipeptídicia, que estará voltada para a interface ar/água. No entanto, ao menos até 30mN/m, nenhuma alteração conformacional foi detectada, como revelada pelos espectros de infravermelho. Filmes LB mistos de DMPA e as duas formas de fosfatase alcalina revelaram que o empacotamento máximo de proteína depende da presença da âncora GFI. Dados usando microgravimetria, atividade enzimática, infravermelho, elipsometria, e microscopia de força atômica mostraram que a adsorção da âncora na interface posiciona o eixo maior do elipsóide, formador da cadeia polipeptídica, paralelamente à matriz lipídica. Na ausência da âncora GFI, o posicionamento da cadeia polipeptídica na interface é aleatório, e para um alto grau de empacotamento, as interações entre os resíduos de aminoácidos intermoleculares favorecerão o posicionamento do eixo maior do elipsóide em uma posição mais perpendicular à interface lipídicaNão const