Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target
<i>Cryptococcus</i> remains a leading cause of invasive fungal infections in immunocompromised people. Resistance to azole drugs has imposed a further challenge to the effective treatment of such infections. In this study, the functional expression of full-length hexahistidine-tagged <...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-11-01
|
| Series: | Journal of Fungi |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2309-608X/8/12/1256 |
| _version_ | 1797456893974675456 |
|---|---|
| author | Yasmeen N. Ruma Mikhail V. Keniya Brian C. Monk |
| author_facet | Yasmeen N. Ruma Mikhail V. Keniya Brian C. Monk |
| author_sort | Yasmeen N. Ruma |
| collection | DOAJ |
| description | <i>Cryptococcus</i> remains a leading cause of invasive fungal infections in immunocompromised people. Resistance to azole drugs has imposed a further challenge to the effective treatment of such infections. In this study, the functional expression of full-length hexahistidine-tagged <i>Cryptococcus neoformans</i> CYP51 (CnCYP51-6×His), with or without its cognate hexahistidine-tagged NADPH-cytochrome P450 reductase (CnCPR-6×His), in a <i>Saccharomyces cerevisiae</i> host system has been used to characterise these enzymes. The heterologous expression of CnCYP51-6×His complemented deletion of the host CYP51 and conferred increased susceptibility to both short-tailed and long-tailed azole drugs. In addition, co-expression of CnCPR-6×His decreased susceptibility 2- to 4-fold for short-tailed but not long-tailed azoles. Type 2 binding of azoles to CnCYP51-6×His and assay of NADPH cytochrome P450 reductase activity confirmed that the heterologously expressed CnCYP51 and CnCPR are functional. The constructs have potential as screening tools and use in structure-directed antifungal discovery. |
| first_indexed | 2024-03-09T16:14:22Z |
| format | Article |
| id | doaj.art-e9dbbfb0dbc14458adad81c0f3c0b9ba |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T16:14:22Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-e9dbbfb0dbc14458adad81c0f3c0b9ba2023-11-24T15:59:22ZengMDPI AGJournal of Fungi2309-608X2022-11-01812125610.3390/jof8121256Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug TargetYasmeen N. Ruma0Mikhail V. Keniya1Brian C. Monk2Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin 9016, New ZealandSir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin 9016, New ZealandSir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin 9016, New Zealand<i>Cryptococcus</i> remains a leading cause of invasive fungal infections in immunocompromised people. Resistance to azole drugs has imposed a further challenge to the effective treatment of such infections. In this study, the functional expression of full-length hexahistidine-tagged <i>Cryptococcus neoformans</i> CYP51 (CnCYP51-6×His), with or without its cognate hexahistidine-tagged NADPH-cytochrome P450 reductase (CnCPR-6×His), in a <i>Saccharomyces cerevisiae</i> host system has been used to characterise these enzymes. The heterologous expression of CnCYP51-6×His complemented deletion of the host CYP51 and conferred increased susceptibility to both short-tailed and long-tailed azole drugs. In addition, co-expression of CnCPR-6×His decreased susceptibility 2- to 4-fold for short-tailed but not long-tailed azoles. Type 2 binding of azoles to CnCYP51-6×His and assay of NADPH cytochrome P450 reductase activity confirmed that the heterologously expressed CnCYP51 and CnCPR are functional. The constructs have potential as screening tools and use in structure-directed antifungal discovery.https://www.mdpi.com/2309-608X/8/12/1256<i>Cryptococcus neoformans</i>CYP51cognate reductaseantifungal resistance<i>Saccharomyces cerevisiae</i> expression systemscreening tool |
| spellingShingle | Yasmeen N. Ruma Mikhail V. Keniya Brian C. Monk Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target Journal of Fungi <i>Cryptococcus neoformans</i> CYP51 cognate reductase antifungal resistance <i>Saccharomyces cerevisiae</i> expression system screening tool |
| title | Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target |
| title_full | Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target |
| title_fullStr | Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target |
| title_full_unstemmed | Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target |
| title_short | Exploring <i>Cryptococcus neoformans</i> CYP51 and Its Cognate Reductase as a Drug Target |
| title_sort | exploring i cryptococcus neoformans i cyp51 and its cognate reductase as a drug target |
| topic | <i>Cryptococcus neoformans</i> CYP51 cognate reductase antifungal resistance <i>Saccharomyces cerevisiae</i> expression system screening tool |
| url | https://www.mdpi.com/2309-608X/8/12/1256 |
| work_keys_str_mv | AT yasmeennruma exploringicryptococcusneoformansicyp51anditscognatereductaseasadrugtarget AT mikhailvkeniya exploringicryptococcusneoformansicyp51anditscognatereductaseasadrugtarget AT briancmonk exploringicryptococcusneoformansicyp51anditscognatereductaseasadrugtarget |