Surface density as a significant parameter for the enzymatic activity of two forms of alkaline phosphatase immobilized on phospholipid Langmuir–Blodgett films

Caseli, Luciano; Furriel, Rosa dos Prazeres Melo; Andrade, José Fernando de; Leone, Francisco de Assis; Zaniquelli, Maria Elisabete Darbello

doi:10.1016/j.jcis.2004.01.081

Cited by 40 publications

(59 citation statements)

References 53 publications

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“…In this sense, to rebuild the surface structure with a high mechanical stability, the conformational and orientation changes must not be highly restricted. In our case, the presence of a phospholipid (hydrophobic anchor) covalently bound to the polypeptidic moiety of the protein gives the GPI-anchored alkaline phosphatase extremely peculiar surface properties, with the GPI-anchor driving the processes of adsorption, orientation, 10,25,[46][47][48] and packing 9 and, as stated by the present work, also influencing the surface elasticity properties. To the best of our knowledge, this is the first report concerning the elasticity properties of a GPI-anchored protein.…”

Section: Elasticity Measurementsmentioning

confidence: 49%

“…Alkaline phosphatase has an ellipsoidal shape, 43 and, according to a previous work, 10 its orientation at the interface (major axis parallel or perpendicular to the interface) is strongly influenced by the presence or not of the hydrophobic anchor. Therefore, the higher or lower capability of the enzyme molecules to pack at the interface, as well as the protein solvation 20,[44][45] may lead to the different velocities of adjustments of the steady state.…”

Section: Elasticity Measurementsmentioning

confidence: 84%

“…9 However, its heterogeneous catalysis differs significantly from that of DSAP. 10 These two enzyme forms provide a very interesting possibility for comparative studies at air/water interfaces, in order to verify the effect of the hydrophobic anchor on the surface properties.…”

Section: Introductionmentioning

confidence: 99%

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Adsorption kinetics and dilatational rheological studies for the soluble and anchored forms of alkaline phosphatase at the air/water interface

Caseli¹,

Masui²,

Furriel³

et al. 2005

J. Braz. Chem. Soc.

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Este trabalho apresenta aspectos de equilíbrio e dinâmicos da adsorção na interface ar/líquido de duas formas de fosfatase alcalina de placa óssea de ratos: DSAP, solubilizada com tensoativo não-iônico (C 12 E 9 ), contendo uma âncora de glicosilfosfatidilinositol (GPI), e PLSAP, com a porção hidrofóbica da âncora clivada por fosfolipase-C. A tensão superficial dinâmica, γ dyn , e o módulo de elasticidade superficial dilatacional, ε, foram determinados para soluções de PLSAP, DSAP e C 12 E 9 pelo método de oscilação harmônica e análise do formato da gota eixo-simétrica. Cinéticas de adsorção revelaram que DSAP adsorve trinta vezes mais rapidamente que PLSAP, apresentando um mínimo, e, para PLSAP, a tensão superficial cai continuamente. Para o sistema DSAP/C 12 E 9 , ε atinge um máximo na concentração crítica de agregação (CAC), mas para PLSAP, ε diminui continuamente com a concentração. Soluções de C 12 E 9 apresentam ε mais elevados, decrescentes com a concentração. Um modelo, baseado na influência da âncora GPI, é proposto para explicar os resultados obtidos.This work presents equilibrium and dynamic aspects for the adsorption at the air/liquid interface of two rat osseous plate alkaline phosphatase forms: DSAP, solubilized by a surfactant, C 12 E 9 , and containing a glycosylphosphatidylinositol (GPI) anchor; and PLSAP, resulting from phospholipase-C cleavage of the hydrophobic portion of the GPI anchor. Dynamic surface tension, γ dyn , and surface elasticity modulus, ε, were determined for PLSAP, DSAP and pure C 12 E 9 solutions using harmonic oscillation and axisymmetric drop shape analysis Adsorption kinetics studies revealed that DSAP adsorbs thirty times faster than PLSAP, presenting a minimum in the curve. For DSAP/ C 12 E 9 mixed system, ε increases with concentration and a maximum appears at the critical aggregation concentration (CAC). For PLSAP, a continuous decreasing with concentration for γ dyn and ε was observed. For pure C 12 E 9 solution , the elasticity modulus increases with concentration and ε values are higher when compared to the mixed system. A model based on the influence of the GPI anchor is proposed.Keywords: dynamic surface tension, adsorption kinetics, dilatational surface elasticity, alkaline phosphatase, axisymmetric drop shape analysis IntroductionSurface elasticity is associated with the ability of a system to establish a new surface tension value due to a timely area variation. Considering that it is a nonequilibrium phenomenon, changes in surface tension occur either as a consequence of a sudden compression/ expansion of the interface, or due to a local concentration variation. Since elasticity values are associated with diffusion processes, adsorption/desorption, as well as the rearrangement of molecules at the interface, they depend on the time scale (or frequency) in which the perturbation is applied. In addition, these processes are influenced by molecular weight, shape and molecular interactions of the species present at the interface.Surface elasticity is directly relate...

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Section: Elasticity Measurementsmentioning

confidence: 49%

Section: Elasticity Measurementsmentioning

confidence: 84%

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Adsorption kinetics and dilatational rheological studies for the soluble and anchored forms of alkaline phosphatase at the air/water interface

Caseli¹,

Masui²,

Furriel³

et al. 2005

J. Braz. Chem. Soc.

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“…The authors observed an increased immobilization rate and subsequent enzyme activity via combination of a weak proximity effect to enzyme carbohydrate moieties, followed by covalent capture mediated by lysine residues of the enzyme. Finally, Caseli et al (2004) used QCM to determine the surface density of alkaline phosphatase adsorbed on to an LB lipid film and subsequent enzyme activity related to the alignment of the immobilized protein molecules.…”

Section: Protein Interactions Protein Adsorptionmentioning

confidence: 99%

A survey of the 2001 to 2005 quartz crystal microbalance biosensor literature: applications of acoustic physics to the analysis of biomolecular interactions

Cooper¹,

Singleton²

2007

J of Molecular Recognition

334

205

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The widespread exploitation of biosensors in the analysis of molecular recognition has its origins in the mid-1990s following the release of commercial systems based on surface plasmon resonance (SPR). More recently, platforms based on piezoelectric acoustic sensors (principally 'bulk acoustic wave' (BAW), 'thickness shear mode' (TSM) sensors or 'quartz crystal microbalances' (QCM)), have been released that are driving the publication of a large number of papers analysing binding specificities, affinities, kinetics and conformational changes associated with a molecular recognition event. This article highlights salient theoretical and practical aspects of the technologies that underpin acoustic analysis, then reviews exemplary papers in key application areas involving small molecular weight ligands, carbohydrates, proteins, nucleic acids, viruses, bacteria, cells and lipidic and polymeric interfaces. Key differentiators between optical and acoustic sensing modalities are also reviewed.

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“…Neste contexto, a geometria da molécula da enzima imobilizada é importante não só para a sua conformação para as funções biológicas, mas também para o posicionamento certo do sítio catalítico no substrato. A desnaturação de enzimas em matrizes sólidas é comum 58,59 . Sua atividade pode ser mais alta 60 Ressalta-se que na formação dos filmes LB, a enzima foi injetada na subfase, sob uma monocamada de DPPG pré-formada.…”

Section: Atividade Enzimáticaunclassified

Estudo da interação da peroxidase de raiz forte em interfaces nanoestruturadas

Schmidt¹

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Surface density as a significant parameter for the enzymatic activity of two forms of alkaline phosphatase immobilized on phospholipid Langmuir–Blodgett films

Cited by 40 publications

References 53 publications

Adsorption kinetics and dilatational rheological studies for the soluble and anchored forms of alkaline phosphatase at the air/water interface

Adsorption kinetics and dilatational rheological studies for the soluble and anchored forms of alkaline phosphatase at the air/water interface

A survey of the 2001 to 2005 quartz crystal microbalance biosensor literature: applications of acoustic physics to the analysis of biomolecular interactions

Estudo da interação da peroxidase de raiz forte em interfaces nanoestruturadas

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