Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence
The inhibition of recombinant CpLIP2 lipase/acyltransferase from <i>Candida parapsiolosis</i> was considered a key model for novel antifungal drug discovery and a potential therapeutic target for candidiasis. Lipases have identified recently as potent virulence factors in <i>C. par...
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MDPI AG
2019-08-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/24/16/2888 |
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| author | Ruba Nasri Luc P R Bidel Nathalie Rugani Véronique Perrier Frédéric Carrière Eric Dubreucq Christian Jay-Allemand |
| author_facet | Ruba Nasri Luc P R Bidel Nathalie Rugani Véronique Perrier Frédéric Carrière Eric Dubreucq Christian Jay-Allemand |
| author_sort | Ruba Nasri |
| collection | DOAJ |
| description | The inhibition of recombinant CpLIP2 lipase/acyltransferase from <i>Candida parapsiolosis</i> was considered a key model for novel antifungal drug discovery and a potential therapeutic target for candidiasis. Lipases have identified recently as potent virulence factors in <i>C. parapsilosis</i> and some other yeasts. The inhibition effects of orlistat and four flavonols (galangin, kaempferol, quercetin and myricetin) characterized by an increasing degree of hydroxylation in B-ring, were investigated using ethyl oleate hydrolysis as the model reaction. Orlistat and kaempferol (14 µM) strongly inhibited CpLIP2 catalytic activity within 1 min of pre-incubation, by 90% and 80%, respectively. The relative potency of flavonols as inhibitors was: kaempferol > quercetin > myricetin > galangin. The results suggested that orlistat bound to the catalytic site while kaempferol interacted with W294 on the protein lid. A static mechanism of interactions between flavonols and CpLIP2 lipase was confirmed by fluorescence quenching analyses, indicating that the interactions were mainly driven by hydrophobic bonds and electrostatic forces. From the Lehrer equation, fractions of tryptophan accessibility to the quencher were evaluated, and a relationship with the calculated number of binding sites was suggested. |
| first_indexed | 2024-12-13T03:16:03Z |
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| id | doaj.art-6ea1987091394a80af39008163767e84 |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-13T03:16:03Z |
| publishDate | 2019-08-01 |
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| series | Molecules |
| spelling | doaj.art-6ea1987091394a80af39008163767e842022-12-22T00:01:29ZengMDPI AGMolecules1420-30492019-08-012416288810.3390/molecules24162888molecules24162888Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of FluorescenceRuba Nasri0Luc P R Bidel1Nathalie Rugani2Véronique Perrier3Frédéric Carrière4Eric Dubreucq5Christian Jay-Allemand6UMR 1208 IATE, Université de Montpellier, 34095 Montpellier, FranceUMR 1334 AGAP, INRA, Place Viala, 34060 Montpellier, FranceDépartement Bio-MV, Université de Montpellier, 34095 Montpellier, FranceUMR 1208 IATE, Université de Montpellier, 34095 Montpellier, FranceUMR 7281 BIP, Aix Marseille Université, CNRS, 31 chemin Joseph Aiguier, 13402 Marseille cedex 09, FranceUMR 1208 IATE, Montpellier SupAgro, Place Viala, 34060 Montpellier, FranceUMR 1208 IATE, Université de Montpellier, 34095 Montpellier, FranceThe inhibition of recombinant CpLIP2 lipase/acyltransferase from <i>Candida parapsiolosis</i> was considered a key model for novel antifungal drug discovery and a potential therapeutic target for candidiasis. Lipases have identified recently as potent virulence factors in <i>C. parapsilosis</i> and some other yeasts. The inhibition effects of orlistat and four flavonols (galangin, kaempferol, quercetin and myricetin) characterized by an increasing degree of hydroxylation in B-ring, were investigated using ethyl oleate hydrolysis as the model reaction. Orlistat and kaempferol (14 µM) strongly inhibited CpLIP2 catalytic activity within 1 min of pre-incubation, by 90% and 80%, respectively. The relative potency of flavonols as inhibitors was: kaempferol > quercetin > myricetin > galangin. The results suggested that orlistat bound to the catalytic site while kaempferol interacted with W294 on the protein lid. A static mechanism of interactions between flavonols and CpLIP2 lipase was confirmed by fluorescence quenching analyses, indicating that the interactions were mainly driven by hydrophobic bonds and electrostatic forces. From the Lehrer equation, fractions of tryptophan accessibility to the quencher were evaluated, and a relationship with the calculated number of binding sites was suggested.https://www.mdpi.com/1420-3049/24/16/2888recombinant CpLIP2 lipaseflavonolsorlistatfluorescence quenchinginhibitionhydroxylationtryptophan residuesdocking |
| spellingShingle | Ruba Nasri Luc P R Bidel Nathalie Rugani Véronique Perrier Frédéric Carrière Eric Dubreucq Christian Jay-Allemand Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence Molecules recombinant CpLIP2 lipase flavonols orlistat fluorescence quenching inhibition hydroxylation tryptophan residues docking |
| title | Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence |
| title_full | Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence |
| title_fullStr | Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence |
| title_full_unstemmed | Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence |
| title_short | Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence |
| title_sort | inhibition of cplip2 lipase hydrolytic activity by four flavonols galangin kaempferol quercetin myricetin compared to orlistat and their binding mechanisms studied by quenching of fluorescence |
| topic | recombinant CpLIP2 lipase flavonols orlistat fluorescence quenching inhibition hydroxylation tryptophan residues docking |
| url | https://www.mdpi.com/1420-3049/24/16/2888 |
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