Relationships between conformational changes and antimicrobial activity of lysozyme upon reduction of its disulfide bonds

Touch, Visalsok; Hayakawa, Shigeru; Saitoh, Koichi

doi:10.1016/s0308-8146(03)00252-8

Cited by 53 publications

(44 citation statements)

References 36 publications

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“…The fully unfolded lysozyme not only had a longer k max wavelength, but also had a higher emission intensity than the native and urea-treated lysozymes. The results confirmed that the disulfide bonds are critical for stabilizing lysozymes (Touch et al 2004). …”

Section: Lysozyme Was Not Unfolded By Urea Alonesupporting

confidence: 58%

“…However, the presence of DTT results in very high absorbance that masks the protein ellipticity and thus hinders the determination of unfolding by circular dichroism (Panda and Horowitz 2000). So, only the intrinsic fluorescence spectrum of tryptophan has been adopted to represent the extent of protein denaturation (Touch et al 2004). When proteins are exposed to a chaotropic reagent, tryptophans in the inner hydrophobic moiety are exposed to the bulk solution, and a red shift in the wavelength at the maximum emission intensity is often observed (Copeland 1993;Ladokhin 2000).…”

Section: Introductionmentioning

confidence: 99%

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Refolding of partially and fully denatured lysozymes

2007

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Lysozyme refolding with high yields sometimes results from incomplete denaturation. Dithiothreitol (DTT) is a reductant commonly used to reduce and unfold disulfide-stabilized lysozymes. Through the use of fluorescence spectroscopy to access the extent of denaturation, we found that the rate and extent of denaturation highly depended on the concentration of DTT. Further, the denaturation exhibited a two-phase transition at a high DTT concentration with DTT at >100 mM and long denaturation time (>24 h) being needed for complete denaturation. A low DTT concentration and a short denaturation time resulted in fast refolding with high activity recovery, while a high DTT concentration and a long denaturation time resulted in slow refolding with low activity recovery. Hence, the renaturation of disulfide-containing lysozyme was highly affected by the extent of denaturation.

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Section: Lysozyme Was Not Unfolded By Urea Alonesupporting

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Refolding of partially and fully denatured lysozymes

2007

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“…In addition, Lz partially unfolded by heat treatment proved to exert catalytically independent antimicrobial action against both G+ and G-bacteria [24]. The enhancement of the bactericidal activity of lysozyme against Gram-negative bacteria was observed also after partial reduction of its disulfide bonds [71]. These results were interpreted in terms of the altered protein conformation minimizing the energetic cost of diffusion through the outer membrane of G-bacteria and enhanced insertion into the inner membrane.…”

Section: Discussionmentioning

confidence: 92%

Modulation of physiological and pathological activities of lysozyme by biological membranes

Trusova

2012

Cellular and Molecular Biology Letters

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Abstract:The molecular details of interactions between lipid membranes and lysozyme (Lz), a small polycationic protein with a wide range of biological activities, have long been the focus of numerous studies. The biological consequences of this process are considered to embrace at least two aspects: i) correlation between antimicrobial and membranotropic properties of this protein, and ii) lipid-mediated Lz amyloidogenesis. The mechanisms underlying the lipid-assisted protein fibrillogenesis and membrane disruption exerted by Lz in bacterial cells are believed to be similar. The present investigation was undertaken to gain further insight into Lz-lipid interactions and explore the routes by which Lz exerts its antimicrobial and amyloidogenic actions. Binding and Förster resonance energy transfer studies revealed that upon increasing the content of anionic lipids in lipid vesicles, Lz forms aggregates in a membrane environment. Total internal reflection fluorescence microscopy and pyrene excimerization reaction were employed to study the effect of Lz on the structural and dynamic properties of lipid bilayers. It was found that Lz induces lipid demixing and reduction of bilayer free volume, the magnitude of this effect being much more pronounced for oligomeric protein.

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“…The reduction time of 1.5 h showed the highest antimicrobial activity against S. enteritidis IFO3313. In cLz, the exposure of hydrophobic region which is buried in the interior of the compact lysozyme molecule was reported after it was reduced by DTT (Touch et al, 2004). Increasing in exposed hydrophobic region is an important factor of binding affinity which can expand the spectrum of Lz antimicrobial activity against gram-negative bacteria, containing of the lipopolysaccharide at the outer membrane.…”

Section: Resultsmentioning

confidence: 99%

“…Many researchers attempt to enhance the antimicrobial activity of Lz against IUFoST 2006DOI: 10.1051 gram-negative bacteria by using chemical or physical treatment to disrupt the bacteria membrane (Vannini et al, 2004), combining with other antimicrobial or/and chemical substances (Facon, 1996;Boland et al, 2003;Branen et al, 2004) and modifying the structure of Lz (Ibrahim et al, 1992;Liu et al, 2000;Mine et al, 2004;Hunter et al, 2005). Recently, reduction of Lz's disulfide bonds, exposing the hydrophobic surface of Lz, was introduced to improve the antimicrobial action of Lz against S. enteritidis (Touch et al, 2004) Most of the reports on antimicrobial activity are about chicken Lz (cLz). Duck Lz (dLz) shows some differences in amino acid sequence.…”

Section: Introductionmentioning

confidence: 99%

Antimicrobial Activity of Duck Egg Lysozyme against Salmonella enteritidis

Hayakawa¹,

Naknukool²,

Uno³

et al. 2006

13th World Congress of Food Science &Amp; Technology

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The objective of the study was to investigate antimicrobial activity of native and reduced forms of duck lysozyme (dLz) against Salmonella enteritidis. Moreover, antimicrobial activity of dLz was compared with chicken lysozyme (cLz). The purified dLz was reduced with dithiothreitol. Free SH groups of reduced dLz were trapped with Combination of lactoferrin (0.1 mg/ml) and reduced dLz (0.1 mg/ml) showed a synergistic effect. In all conditions, dLz showed higher activity than cLz in both native and reduced forms. The result of the study indicated that reduced dLz trends to be a more efficient antimicrobial agent against S. enteritidis. To achieve the highest sufficiency, the essential roles of food components and incubation temperature should be concerned in reduced dLz application.

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Relationships between conformational changes and antimicrobial activity of lysozyme upon reduction of its disulfide bonds

Cited by 53 publications

References 36 publications

Refolding of partially and fully denatured lysozymes

Refolding of partially and fully denatured lysozymes

Modulation of physiological and pathological activities of lysozyme by biological membranes

Antimicrobial Activity of Duck Egg Lysozyme against Salmonella enteritidis

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