Alliin is a suicide substrate of<i>Citrobacter freundii</i>methionine γ-lyase: structural bases of inactivation of the enzyme

Morozova, Elena A.; Revtovich, Svetlana V.; Anufrieva, Natalya V.; Kulikova, Vitalia V.; Nikulin, Alexei; Demidkina, Tatyana V.

doi:10.1107/s1399004714020938

Cited by 18 publications

(22 citation statements)

References 40 publications

(57 reference statements)

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“…Therefore, it is one of the hotspot research areas in the field of cancer to explore new therapy for regulating the cellular mechanisms and inducing apoptosis to treat tumors and more and more studies are focusing on natural products. Allicin is a sulfur compound extracted from garlic bulbs [11,39]. The surveys of epidemiology show that allicin has an inhibition effect on many malignant tumors.…”

Section: Discussionmentioning

confidence: 99%

Role of p38 MAPK activation and mitochondrial cytochrome-c release in allicin-induced apoptosis in SK-N-SH cells

Zhuang

Chi

2016

Anti-Cancer Drugs

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Here, we investigate the apoptotic effect of allicin, the predominant component of freshly crushed garlic, on neuroblastoma cells. In this paper, the authors have first assessed the effect of allicin on human neuroblastoma SK-N-SH cells and then investigated the underlying mechanism. The results indicate that allicin suppresses SK-N-SH cell growth in a dose-dependent and time-dependent manner and that 5 μmol/l of allicin leads to a significant increase in apoptotic rate with annexin-V/PI double staining. Western blot analysis shows that treatment with allicin-induced apoptosis through activation of caspases-3 and 9. Phosphorylation of p38 MAPK contributes to allicin-induced apoptosis upstream of caspase activation. Using p38 MAPK inhibitor, the authors discovered that p38 MAPK activation subsequently induces the release of cytochrome-c from mitochondria into the cytosol. Taken together, the results demonstrate that allicin can activate the p38 MAPK pathway, which leads to mitochondrial release of cytochrome-c, thus inducing SK-N-SH cell apoptosis. Overall, this study suggests that allicin may be used as one of the novel pharmacological treatment strategies in neuroblastoma.

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Section: Discussionmentioning

confidence: 99%

Role of p38 MAPK activation and mitochondrial cytochrome-c release in allicin-induced apoptosis in SK-N-SH cells

Zhuang

Chi

2016

Anti-Cancer Drugs

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“…Recently, we have showed that PLP‐dependent MGL (EC 4.4.1.11) from Citrobacter freundii catalyzes the β‐elimination reaction of alliin and other S‐substituted l ‐cysteine sulfoxides to form allicin or S‐alkylthiosulfinates:

…”

Section: Introductionmentioning

confidence: 99%

“…C. freundii MGL contains cysteine 115 residue in the active site . To avoid the observed 31% inactivation of the enzyme in the β‐elimination reaction of alliin , we prepared the mutant form of C. freundii MGL with the replacement of cysteine 115 by histidine. The replacement allowed us to obtain an effective catalyst for the production of thiosulfinates .…”

Section: Introductionmentioning

confidence: 99%

Mutant formC115HofClostridium sporogenesmethionine γ‐lyase efficiently cleavesS‐Alk(en)yl‐l‐cysteine sulfoxides to antibacterial thiosulfinates

et al. 2016

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Pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) catalyzes the β-elimination reaction of S-alk(en)yl-l-cysteine sulfoxides to thiosulfinates, which possess antimicrobial activity. Partial inactivation of the enzyme in the course of the reaction occurs due to oxidation of active site cysteine 115 conserved in bacterial MGLs. In this work, the C115H mutant form of Clostridium sporogenes MGL was prepared and the steady-state kinetic parameters of the enzyme were determined. The substitution results in an increase in the catalytic efficiency of the mutant form towards S-substituted l-cysteine sulfoxides compared to the wild type enzyme. We used a sulfoxide/enzyme system to generate antibacterial activity in situ. Two-component systems composed of the mutant enzyme and three S-substituted l-cysteine sulfoxides were demonstrated to be effective against Gram-positive and Gram-negative bacteria and three clinical isolates from mice. © 2016 IUBMB Life, 68(10):830-835, 2016.

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“…In C. freundii, among the seven cysteine residues present in the amino acid sequence, two are highly reactive to modification by DTNB. According to crystallographic data, C4 and C115 are the most plausible candidates . In C. novyi MGL, we determined that one SH group reacts rapidly with DTNB under native conditions and three SH groups react with DTNB in 6 M guanidine hydrochloride, as expected on the basis of the amino acid sequence.…”

Section: Resultsmentioning

confidence: 81%

“…Recently, we provided evidence for the cytotoxicity of MGL from C. sporogenes, C. tetani, and Citrobacter freundii MGL using K562, PC-3, LnCap, MCF7, SKOV-3, and L5178y tumor cell lines (9). We also reported the capability of MGL to catalyze the formation of the potent antibacterial agent allicin or Salkylthiosulfinates from substrate analogues, such as alliin and S-substituted L-cysteine sulfoxides (10,11).…”

Section: Introductionmentioning

confidence: 94%

Gene cloning, characterization, and cytotoxic activity of methionine γ‐lyase from Clostridium novyi

et al. 2017

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The exploitation of methionine-depleting enzyme methionine γ-lyase (MGL) is a promising strategy against specific cancer cells that are strongly dependent on methionine. To identify MGL from different sources with high catalytic activity and efficient anticancer action, we have expressed and characterized MGL from Clostridium novyi and compared its catalytic efficiency with the previously studied MGL from Citrobacter freundii. The purified recombinant MGL exhibits k and k /K for methionine γ-elimination reaction that are 2.4- and 1.36-fold higher than C. freundii enzyme, respectively, whereas absorption, fluorescence, and circular dichroism spectra are very similar, as expected on the basis of 87% sequence identity and high conservation of active site residues. The reactivity of cysteine residues with DTNB and iodoacetamide was investigated as well as the impact of their chemical modification on catalytic activity. This information is relevant because for increasing bioavailability and reducing immunogenity, MGL should be decorated with polyethylene glycol (PEG). It was found that Cys118 is a faster reacting residue, which results in a significant decrease in the γ-elimination activity. Thus, the protection of Cys118 before conjugation with cysteine-reacting PEG represents a valuable strategy to preserve MGL activity. The anticancer action of C. novyi MGL, evaluated in vitro against prostate (PC-3), chronic myelogenous leucemia (K562), and breast (MDA-MB-231 and MCF7) cancer cells, exhibits IC of 1.3 U mL , 4.4 U mL , 1.2 U mL , and 3.4 U mL , respectively. A higher cytotoxicity of C. novyi MGL was found against cancer cells with respect to C. freundii MGL, with the exception of PC-3, where a lower cytotoxicity was observed. © 2017 IUBMB Life, 69(9):668-676, 2017.

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Alliin is a suicide substrate ofCitrobacter freundiimethionine γ-lyase: structural bases of inactivation of the enzyme

Cited by 18 publications

References 40 publications

Role of p38 MAPK activation and mitochondrial cytochrome-c release in allicin-induced apoptosis in SK-N-SH cells

Role of p38 MAPK activation and mitochondrial cytochrome-c release in allicin-induced apoptosis in SK-N-SH cells

Mutant formC115HofClostridium sporogenesmethionine γ‐lyase efficiently cleavesS‐Alk(en)yl‐l‐cysteine sulfoxides to antibacterial thiosulfinates

Gene cloning, characterization, and cytotoxic activity of methionine γ‐lyase from Clostridium novyi

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