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1.
Colloids Surf B Biointerfaces ; 103: 416-21, 2013 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-23261561

RESUMO

Competitive adsorption is a general problem both in polymer and in biological systems. The equilibrium composition at a surface in contact either with polymer solutions or biological fluids depends on the competition between all the surface active material present in the medium. Such competition is particularly important in cell membranes where membrane proteins generated on ribosomes have to incorporate in the cell. Here we use fluovideo microscopy to study the competition for adsorption at the air/water interface between the enzyme glucose oxidase (GOx) and fluid monolayers of pentadecanoic acid (PDA). Although water soluble, GOx has a strong affinity for the air/water interface. We show that under certain conditions it inserts in the monolayer and causes a contraction of the Langmuir film and the formation of condensed domains. When exposed to a heterogeneous surface it is inserted in the less dense regions. Its crystallization leads to the deformation of the condensed domains followed by the destruction of their initial shape. By compressing the layer the protein is not removed from the interface where it eventually forms three-dimensional structures.


Assuntos
Ácidos Graxos/metabolismo , Glucose Oxidase/metabolismo , Ácido Mirístico/metabolismo , Adsorção , Aspergillus niger/enzimologia , Microscopia de Fluorescência , Pressão , Temperatura , Xantenos/metabolismo
3.
J Biochem Biophys Methods ; 62(1): 51-62, 2005 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-15656943

RESUMO

The recognition of DNA is the first and most important condition for biological applications, including transcription and translation regulators and DNA sensors. For this purpose, we have developed few systems where we were able to immobilize long double-stranded DNA (dsDNA) successfully to the surfaces of different solid substrates. To achieve this, we have chosen polystyrene beads and standard Langmuir-Blodgett monolayer of Zn-arachidate. In the first attempt, variant of T7 DNA containing one strong promoter A1 for Escherichia coli RNA polymerase was immobilised on uniform polystyrene microspheres (0.31 microm diameter) by covalent grafting. In the latter case, Zn(II) is bound to arachidic acid through charge neutralization. Since tetrahedral Zn(II) participates in DNA recognition through coordination, we have been able to layer DNA over the Zn-arachidate monolayer. The successful immobilization of DNAs on these different substrates was visualized under fluorescence microscope. These immobilized DNAs were used as a template to study in vitro transcription reaction and thus we introduce a new strategy for the study of transcription in heterogeneous phase.


Assuntos
Bacteriófago T7/genética , Sondas de DNA/química , DNA Viral/química , Membranas Artificiais , Transcrição Gênica , Adsorção , Materiais Revestidos Biocompatíveis/química , Sondas de DNA/genética , DNA Viral/genética , DNA Viral/ultraestrutura , RNA Polimerases Dirigidas por DNA/química , Ácidos Eicosanoicos/química , Microesferas
4.
Langmuir ; 20(14): 5891-6, 2004 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-16459606

RESUMO

Here, we report a system we have developed where long double-stranded DNAs (dsDNAs) are immobilized on a monolayer of Zn-arachidate. We have applied the Langmuir-Blodgett technique to form the monolayer of Zn-arachidate where Zn(II) is bound to arachidic acid through charge neutralization. Because tetrahedral Zn(II) participates in DNA recognition through coordination, we have been able to layer DNA over the Zn-arachidate monolayer. The DNA layer shows a typical compression and expansion cycle in a concentration-dependent fashion. Interestingly, the DNA monolayer is available for enzymatic degradation by DNaseI. The detection of DNA and its accessibility towards biological reaction is demonstrated by imaging through fluorescence microscopy. The conformation of the DNA, immobilized on the monolayer, was studied with the help of atomic force microscopy (AFM). We observed that the dsDNAs were aligned in a stretched manner on the surface. To investigate further, we also demonstrate here that the small single-stranded DNA (ssDNA) immobilized on the air-water interface can act as a target molecule for the complementary ssDNA present in the subphase. The study of DNA hybridization done with the help of fluorescence spectroscopy clearly supports the AFM characterization.


Assuntos
DNA Bacteriano/química , Desoxirribonuclease I/química , Membranas Artificiais , Ácidos Eicosanoicos/química , Vidro/química , Propriedades de Superfície , Zinco/química
5.
Chem Commun (Camb) ; (10): 1186-7, 2003 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-12778727

RESUMO

A variety of groups like a Fischer carbene complex, an N-hydroxysuccinimide or a ferrocene derivative have been grafted by ruthenium-catalyzed cross-metathesis reaction with terminal alkene groups on monolayer-protected gold clusters as a mild and convenient strategy to anchor functional molecules.

6.
Phys Rev Lett ; 90(12): 126104, 2003 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-12688889

RESUMO

We have measured the shear viscosity in the liquid phase of several Langmuir monolayers with an accuracy better than 30%. The method is based on the optical monitoring of the Brownian diffusion of submicron latex spheres floating at the air-water interface. The values are between 1 and 11x10(-10) N s m(-1), which is 10 to 100 times lower than previous data on similar systems. For N-palmitoyl-6-n-penicillanic acid and L-alpha-dipalmitoylphosphatidylcholine, the variation of the shear viscosity with surface density agrees with a classical free area model, whereas for pentadecanoic acid we observe a compensation effect.

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