Covalent conjugation of tetrameric bovine liver catalase to liposome membranes for stabilization of the enzyme tertiary and quaternary structures

Yoshimoto, Makoto; Sakamoto, Hideyuki; Shirakami, Hiroshi

doi:10.1016/j.colsurfb.2008.11.028

Cited by 30 publications

(16 citation statements)

References 26 publications

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“…Within such nanoenvironments, the motions of water molecules are so much restricted that they do not crystallize even at temperatures close to the absolute zero but instead, transition into a highly viscous state, stabilizing the protein entities by hyper-increasing its rotational, translational and vibrational viscosity [8,22,64,83,124]. Confining multimeric proteins within liposomes have been proposed as a general strategy to stabilize such protein entities [125]. Additionally, reverse micelles (closed, almost spherical aggregates of surfactant molecules (e.g.…”

Section: Stabilization Via Physical Containmentmentioning

confidence: 99%

Structural and functional stabilization of protein entities: state-of-the-art

Balcão

Vila

2015

Advanced Drug Delivery Reviews

180

104

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Within the context of biomedicine and pharmaceutical sciences, the issue of (therapeutic) protein stabilization assumes particular relevance. Stabilization of protein and protein-like molecules translates into preservation of both structure and functionality during storage and/or targeting, and such stabilization is mostly attained through establishment of a thermodynamic equilibrium with the (micro)environment. The basic thermodynamic principles that govern protein structural transitions and the interactions of the protein molecule with its (micro)environment are, therefore, tackled in a systematic fashion. Highlights are given to the major classes of (bio)therapeutic molecules, viz. enzymes, recombinant proteins, (macro)peptides, (monoclonal) antibodies and bacteriophages. Modification of the microenvironment of the biomolecule via multipoint covalent attachment onto a solid surface followed by hydrophilic polymer co-immobilization, or physical containment within nanocarriers, are some of the (latest) strategies discussed aiming at full structural and functional stabilization of said biomolecules.

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Section: Stabilization Via Physical Containmentmentioning

confidence: 99%

Structural and functional stabilization of protein entities: state-of-the-art

Balcão

Vila

2015

Advanced Drug Delivery Reviews

180

104

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“…4 ). The total cholesterol levels were highly correlated (r 2 = tions at 37°C ( 11,12 ), and the >1000-fold higher affi nity of HRP versus catalase for hydrogen peroxide ( 13,14 ). To evaluate the accuracy of the enzymatic method, we compared it with stable isotope dilution GC-MS, the goldstandard method for free cholesterol and cholesterol ester quantifi cation.…”

Section: Comparison Of the Enzymatic And The Gc-ms Assaysmentioning

confidence: 99%

A simple and sensitive enzymatic method for cholesterol quantification in macrophages and foam cells

Robinet

Wang

Hazen

et al. 2010

Journal of Lipid Research

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This article is available online at http://www.jlr.org levels can be separated into two groups: 1 ) analytical methods such as gas-liquid chromatography or liquid chromatography coupled with fl ame ionization or mass spectrometry detection and quantifi cation ( 2-4 ); and 2 ) enzymatic assays, which can be colorimetric ( 5, 6 ) or fl uorometric ( 7 ). However, the chromatographic and mass spectrometry methods require specialized equipment and chemical saponifi cation of cholesterol esters; furthermore, these methods are time consuming and require processing samples one at a time. In contrast, the colorimetric and fl uorometric enzymatic methods are simple, sensitive, and relatively fast assays that can process many samples at once. The principal of a sensitive fl uorometric assay for the measurement of total cholesterol is shown below, with free cholesterol measured by omitting the fi rst esterase step, and cholesterol mass in cholesterol esters calculated as the difference between the total and free cholesterol contents. HRPThe enzymatic method is the principal method used for measuring cholesterol levels in plasma, where cholesterol is solubilized in an aqueous solution through its incorporation in lipoproteins. However, in order to accurately measure the cholesterol content of cells or tissue by this method, one must identify a suitable solvent that allows good recovery of free and esterifi ed cholesterol in solution and that is compatible with the enzymes and substrates. Previously, the use of the enzymatic cholesterol assay for cells and tissues has been limited due to the inadequate quantitative recovery of free cholesterol and cholesterol esters in enzyme compatible solvents; in fact, a methods paper has argued that the enzymatic method cannot be used for this purpose due to poor extraction and solubiliAbstract A precise and sensitive method for measuring cellular free and esterifi ed cholesterol is required in order to perform studies of macrophage cholesterol loading, metabolism, storage, and effl ux. Until now, the use of an enzymatic cholesterol assay, commonly used for aqueous phase plasma cholesterol assays, has not been optimized for use with solid phase samples such as cells, due to ineffi cient solubilization of total cholesterol in enzyme compatible solvents. We present an effi cient solubilization protocol compatible with an enzymatic cholesterol assay that does not require chemical saponifi cation or chromatographic separation. Another issue with enzyme compatible solvents is the presence of endogenous peroxides that interfere with the enzymatic cholesterol assay. We overcame this obstacle by pretreatment of the reaction solution with the enzyme catalase, which consumed endogenous peroxides resulting in reduced background and increased sensitivity in our method. Finally, we demonstrated that this method for cholesterol quantifi cation in macrophages yields results that are comparable to those measured by stable isotope dilution gas chromatography with mass spectrometry detection. In conclusio...

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“…These results can be attributed to a superior enzyme-substrate complex formation conditions upon encapsulation than that offered for the free CAT. These results can be explained regarding the conformational stabilization of the immobilized CAT that may have resulted in a higher accessibility of the substrate to the active sites of the immobilized enzyme [16,38]. Figure 3 presents pH activity profiles of free CAT and e-CAT.…”

Section: Effect Of Encapsulation On Kinetic Parametersmentioning

confidence: 96%

Development of novel delivery system for nanoencapsulation of catalase: formulation, characterization, and in vivo evaluation using oxidative skin injury model

Abdel-Mageed

Fahmy

Shaker

et al. 2018

Artificial Cells, Nanomedicine, and Biotechnology

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One of the main challenges for successful pharmaceutical application of Catalase (CAT) is maintaining its stability. Physical immobilization of CAT through nano-encapsulation was proposed to resolve this challenge. CAT encapsulating niosomes (e-CAT) were prepared using Brij 30, 52, 76, 92, and 97 in the presence of cholesterol (Ch) by thin film hydration method. Niosomes were characterized for encapsulation efficiency % (EE), size, poly-dispersity index (PI), and morphology. Kinetic parameters, pH optimum, thermal stability, and reusability of CAT were determined. The influence of optimized e-CAT dispersion onto thermally injured rat skin was evaluated. Results revealed that encapsulation enhanced CAT catalytic efficiency (V/K). Free CAT and e-CAT had pH optimum at 7.0. e-CAT exhibited improved thermal stability where it retained 50% residual activity at 60 °C. Free CAT lost its activity after three consecutive operational cycles; however, e-CAT retained 60% of its initial activity following 12 cycles. After 24 h of topical application on thermal injury, a significant difference in lesion size was observed with e-CAT compared with the control group. Based on these encouraging results, CAT immobilization demonstrated a promising novel delivery system that enhances its operational stability. In addition, nano-encapsulated CAT can be anticipated to be beneficial in skin oxidative injury.

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Covalent conjugation of tetrameric bovine liver catalase to liposome membranes for stabilization of the enzyme tertiary and quaternary structures

Cited by 30 publications

References 26 publications

Structural and functional stabilization of protein entities: state-of-the-art

Structural and functional stabilization of protein entities: state-of-the-art

A simple and sensitive enzymatic method for cholesterol quantification in macrophages and foam cells

Development of novel delivery system for nanoencapsulation of catalase: formulation, characterization, and in vivo evaluation using oxidative skin injury model

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