Files in this item
Sterol 14-alpha demethylase (CYP51) activity in Leishmania donovani is likely dependent upon cytochrome P450 reductase 1
Item metadata
dc.contributor.author | Tulloch, Lindsay B | |
dc.contributor.author | Tinti, Michele | |
dc.contributor.author | Wall, Richard J | |
dc.contributor.author | Weidt, Stefan K | |
dc.contributor.author | Corpas-Lopez, Victoriano | |
dc.contributor.author | Dey, Gourav | |
dc.contributor.author | Smith, Terry K | |
dc.contributor.author | Fairlamb, Alan H | |
dc.contributor.author | Barrett, Michael P | |
dc.contributor.author | Wyllie, Susan | |
dc.date.accessioned | 2024-07-26T10:30:07Z | |
dc.date.available | 2024-07-26T10:30:07Z | |
dc.date.issued | 2024-07-11 | |
dc.identifier | 305685564 | |
dc.identifier | 1b379a79-1264-40df-81be-2bf15296e954 | |
dc.identifier | 38991025 | |
dc.identifier | 85198307165 | |
dc.identifier.citation | Tulloch , L B , Tinti , M , Wall , R J , Weidt , S K , Corpas-Lopez , V , Dey , G , Smith , T K , Fairlamb , A H , Barrett , M P & Wyllie , S 2024 , ' Sterol 14-alpha demethylase (CYP51) activity in Leishmania donovani is likely dependent upon cytochrome P450 reductase 1 ' , PLoS Pathogens , vol. 20 , no. 7 , e1012382 . https://doi.org/10.1371/journal.ppat.1012382 | en |
dc.identifier.issn | 1553-7366 | |
dc.identifier.other | Jisc: 2130772 | |
dc.identifier.other | pii: PPATHOGENS-D-24-00322 | |
dc.identifier.uri | https://hdl.handle.net/10023/30270 | |
dc.description | Funding: This work was supported by the following Wellcome Trust (https://wellcome.org/) grants: 203134/Z/16/Z (SW and AHF) and 218448/Z/19/Z (SW). LBT, MT, VCL, GD and RW were supported through the grants awarded to SW. MPB was funded by an MRC (https://www.ukri.org/councils/ mrc/) Newton grant: MR/S0196501. SKW is part of the Glasgow University CMVLS research facility. | en |
dc.description.abstract | Liposomal amphotericin B is an important frontline drug for the treatment of visceral leishmaniasis, a neglected disease of poverty. The mechanism of action of amphotericin B (AmB) is thought to involve interaction with ergosterol and other ergostane sterols, resulting in disruption of the integrity and key functions of the plasma membrane. Emergence of clinically refractory isolates of Leishmania donovani and L. infantum is an ongoing issue and knowledge of potential resistance mechanisms can help to alleviate this problem. Here we report the characterisation of four independently selected L. donovani clones that are resistant to AmB. Whole genome sequencing revealed that in three of the moderately resistant clones, resistance was due solely to the deletion of a gene encoding C24-sterol methyltransferase (SMT1). The fourth, hyper-resistant resistant clone (>60-fold) was found to have a 24 bp deletion in both alleles of a gene encoding a putative cytochrome P450 reductase (P450R1). Metabolic profiling indicated these parasites were virtually devoid of ergosterol (0.2% versus 18% of total sterols in wild-type) and had a marked accumulation of 14-methylfecosterol (75% versus 0.1% of total sterols in wild-type) and other 14-alpha methylcholestanes. These are substrates for sterol 14-alpha demethylase (CYP51) suggesting that this enzyme may be a bona fide P450R specifically involved in electron transfer from NADPH to CYP51 during catalysis. Deletion of P450R1 in wild-type cells phenocopied the metabolic changes observed in our AmB hyper-resistant clone as well as in CYP51 nulls. Likewise, addition of a wild type P450R1 gene restored sterol profiles to wild type. Our studies indicate that P450R1 is essential for L. donovani amastigote viability, thus loss of this gene is unlikely to be a driver of clinical resistance. Nevertheless, investigating the mechanisms underpinning AmB resistance in these cells provided insights that refine our understanding of the L. donovani sterol biosynthetic pathway. | |
dc.format.extent | 23 | |
dc.format.extent | 2199611 | |
dc.language.iso | eng | |
dc.relation.ispartof | PLoS Pathogens | en |
dc.subject | NADPH-ferrihemoprotein Reductase - metabolism - genetics | en |
dc.subject | Drug resistance | en |
dc.subject | Amphotericin B - pharmacology | en |
dc.subject | Sterol 14-demethylase - metabolism - genetics | en |
dc.subject | Protozoan proteins - metabolism - genetics | en |
dc.subject | Antiprotozoal agents - pharmacology | en |
dc.subject | Humans | en |
dc.subject | Leishmania donovani - enzymology | en |
dc.subject | Leishmaniasis, visceral - parasitology - drug therapy | en |
dc.subject | Ergosterol - metabolism | en |
dc.subject | QH301 Biology | en |
dc.subject | DAS | en |
dc.subject | SDG 3 - Good Health and Well-being | en |
dc.subject.lcc | QH301 | en |
dc.title | Sterol 14-alpha demethylase (CYP51) activity in Leishmania donovani is likely dependent upon cytochrome P450 reductase 1 | en |
dc.type | Journal article | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.contributor.institution | University of St Andrews. School of Biology | en |
dc.contributor.institution | University of St Andrews. Sir James Mackenzie Institute for Early Diagnosis | en |
dc.identifier.doi | https://doi.org/10.1371/journal.ppat.1012382 | |
dc.description.status | Peer reviewed | en |
This item appears in the following Collection(s)
Items in the St Andrews Research Repository are protected by copyright, with all rights reserved, unless otherwise indicated.