Tryptophan mediated photoreduction of disulfide bond causes unusual fluorescence behaviour of <i>Fusarium solani pisi</i> cutinase

Prompers, Jeanine J.; Hilbers, Cornelis W.; Pepermans, Henri

doi:10.1016/s0014-5793(99)00990-4

Cited by 71 publications

(75 citation statements)

References 32 publications

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“…Disulphide bridges are known to be excellent quenchers of excited-state aromatic residues. However, selective prolonged irradiation of the Trp residue at 295 nm increases its fluorescence quantum yield, in agreement with previous results by Weisenborn et al (1996) and Prompers et al (1999). It was also observed that on irradiation of cutinase with light at 295 nm, free thiol groups are present in cutinase (see Fig.…”

Section: Discussionsupporting

confidence: 91%

“…Because Prompers et al (1999) reported none of the above parameters on which p depends, we can not compare their findings with ours. Also, the concentration of protein used by Prompers et al (500 M cutinase solution) is drastically higher then ours (2, 4, 14, and 17 M), making comparison impossible.…”

Section: Discussioncontrasting

confidence: 57%

“…The probability p for this event is a general physical parameter that is independent of light intensity, beam geometry, and protein concentration. Because of significantly different experimental conditions, our results unfortunately can not be quantitatively compared with results reported by Weisenborn et al (1996) and Prompers et al (1999). The latter paper reports results obtained under severe inner filter effects, the former paper reports results obtained under partial illumination of the sample (i.e., diffusion related effects may distort the observations).…”

Section: Discussionmentioning

confidence: 70%

“…A partial answer on the effect of UV light on the fungal lipase/esterase cutinase from Fusarium solani pisi has been provided by Weisenborn et al (1996) and Prompers et al (1999). Weisenborn et al (1996) reported that the fluorescence quantum yield of cutinase increases with the illumination time of cutinase, and that the short lifetime component, ∼40 ps, observed for Trp before extended illumination is substituted by longer lifetimes on prolonged UV irradiation of cutinase.…”

mentioning

confidence: 99%

“…The UV illumination has been reported to not affect cutinase lipolytic activity, in contradiction with our findings. The disruption by UV light of the disulphide bridge nearby the only Trp residue in cutinase has been correlated by Prompers et al (1999), with the UV excitation of this aromatic residue. Because disulphide bridges are excellent quenchers of the excited state of aromatic residues, once disrupted by UV light, the above-mentioned disulphide bridge could no longer quench the Trp residue.…”

mentioning

confidence: 99%

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High probability of disrupting a disulphide bridge mediated by an endogenous excited tryptophan residue

et al. 2002

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It is well known that ultraviolet (UV) radiation may reduce or even abolish the biological activity of proteins and enzymes. UV light, as a component of sunlight, is illuminating all light-exposed parts of living organisms, partly composed of proteins and enzymes. Although a considerable amount of empirical evidence for UV damage has been compiled, no deeper understanding of this important phenomenon has yet emerged. The present paper presents a detailed analysis of a classical example of UV-induced changes in three-dimensional structure and activity of a model enzyme, cutinase from Fusarium solani pisi. The effect of illumination duration and power has been investigated. A photon-induced mechanism responsible for structural and functional changes is proposed. Tryptophan excitation energy disrupts a neighboring disulphide bridge, which in turn leads to altered biological activity and stability. The loss of the disulphide bridge has a pronounced effect on the fluorescence quantum yield, which has been monitored as a function of illumination power. A general theoretical model for slow two-state chemical exchange is formulated, which allows for calculation of both the mean number of photons involved in the process and the ratio between the quantum yields of the two states. It is clear from the present data that the likelihood for UV damage of proteins is directly proportional to the intensity of the UV radiation. Consistent with the loss of the disulphide bridge, a complex pH-dependent change in the fluorescence lifetimes is observed. Earlier studies in this laboratory indicate that proteins are prone to such UV-induced radiation damage because tryptophan residues typically are located as next spatial neighbors to disulphide bridges. We believe that these observations may have far-reaching implications for protein stability and for assessing the true risks involved in increasing UV radiation loads on living organisms.Keywords: tryptophan fluorescence lifetime; fluorescence quenching; disulphide (disulfide) bridges; photochemical reaction; protein structure damaged by UV light; SS bond disruption; indole; theoretical model Two divergent theories of the mechanisms involved in ultraviolet (UV) inactivation of enzymes have been developed over a period of years. One theory proposes that the random destruction of any amino acid residue causes inactivation (Augenstein and Riley 1964). The second emphasizes the importance of the disruption of a cluster of specific cystine residues (cysteines involved in disulphide bridges; Augenstein and Riley 1964). It was also found that the effective destruction of cystine, tryptophan, tyrosine, and phenylalanine occurs on UV irradiation of proteins (Kazutomo

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Section: Discussionsupporting

confidence: 91%

Section: Discussioncontrasting

confidence: 57%

Section: Discussionmentioning

confidence: 70%

mentioning

confidence: 99%

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confidence: 99%

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High probability of disrupting a disulphide bridge mediated by an endogenous excited tryptophan residue

et al. 2002

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Cutin from Plants

Kolattukudy

2002

Biopolymers Online

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Introduction Historical Outline Occurrence and Ultrastructure of Cutin Isolation of Cutin Depolymerization of Cutin Chemical Depolymerization Enzymatic Depolymerization Monomer Composition of Cutin Structure of Cutin Biosynthesis of Cutin Biosynthesis of the C 16 Family of Cutin Monomers Biosynthesis of the C 18 Family of Cutin Monomers Synthesis of Cutin from Monomers Cutin Biodegradation Cutin Degradation in Plants Degradation of Cutin by Animals Cutin Degradation by Bacteria Fungal Degradation Function of Cutin Material Exchange with the Environment Low‐temperature Adaptation Role of Cutin in the Interaction with Microbes Cutin Required for Proper Development of Plant Organs Potential Commercial Use for Cutin and Cutinase Outlook and Perspectives Patents

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One‐Step Photo Synthesis of Protein–Drug Nanoassemblies for Drug Delivery

Xie

Cao

Mao

et al. 2013

Adv Healthcare Materials

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Upon controlled UV illumination, disulfide bonds in bovine α‐lactalbumin (BLA) are selectively broken, leading to self‐assembly of the BLA and doxorubicin (DOX) molecules into nanoparticles via hydrophobic interactions and intermolecular disulfide bonds. Such protein–drug nanoparticles have synergistic anticancer activity in vitro and tumor‐homing specificity in vivo, which are of great potential for systemic drug delivery in cancer therapy.

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Tryptophan mediated photoreduction of disulfide bond causes unusual fluorescence behaviour of Fusarium solani pisi cutinase

Cited by 71 publications

References 32 publications

High probability of disrupting a disulphide bridge mediated by an endogenous excited tryptophan residue

High probability of disrupting a disulphide bridge mediated by an endogenous excited tryptophan residue

Cutin from Plants

One‐Step Photo Synthesis of Protein–Drug Nanoassemblies for Drug Delivery

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