Proteopedia

9ium

The structure of Candida albicans Cdr1 in milbemycin oxime-inhibited stateThe structure of Candida albicans Cdr1 in milbemycin oxime-inhibited state

Structural highlights

9ium is a 1 chain structure with sequence from Candida albicans SC5314. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:Electron Microscopy, Resolution 3.08Å
Ligands:,
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

CDR1_CANAL Pleiotropic ABC efflux transporter that confers resistance to numerous chemicals including anisomycin, cycloheximide, fluconazole, miconazole, ketoconazole, itriconazole, nystatin, terbinafine, amorolfine, brefeldin A, amphotericin B, fluphenazine, as well as estrogen. Plays a role in farnesol-induced apoptotic process through glutathione efflux activity. Mediates in-to-out translocation of membrane phospholipids including aminophospholipids and thus regulates asymmetric distribution of phosphatidylethanolamine. Exhibits nucleoside triphosphatase activity.[1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] [19] [20] [21] [22] [23] [24] [25] [26] [27] [28] [29] [30] [31] [32]

Publication Abstract from PubMed

In Candida albicans, Cdr1 pumps azole drugs out of the cells to reduce intracellular accumulation at detrimental concentrations, leading to azole-drug resistance. Milbemycin oxime, a veterinary anti-parasitic drug, strongly and specifically inhibits Cdr1. However, how Cdr1 recognizes and exports azole drugs, and how milbemycin oxime inhibits Cdr1 remain unclear. Here, we report three cryo-EM structures of Cdr1 in distinct states: the apo state (Cdr1(Apo)), fluconazole-bound state (Cdr1(Flu)), and milbemycin oxime-inhibited state (Cdr1(Mil)). Both the fluconazole substrate and the milbemycin oxime inhibitor are primarily recognized within the central cavity of Cdr1 through hydrophobic interactions. The fluconazole is suggested to be exported from the binding site into the environment through a lateral pathway driven by TM2, TM5, TM8 and TM11. Our findings uncover the inhibitory mechanism of milbemycin oxime, which inhibits Cdr1 through competition, hindering export, and obstructing substrate entry. These discoveries advance our understanding of Cdr1-mediated azole resistance in C. albicans and provide the foundation for the development of innovative antifungal drugs targeting Cdr1 to combat azole-drug resistance.

Cryo-EM structures of Candida albicans Cdr1 reveal azole-substrate recognition and inhibitor blocking mechanisms.,Peng Y, Lu Y, Sun H, Ma J, Li X, Han X, Fang Z, Tan J, Qiu Y, Qu T, Yin M, Yan Z Nat Commun. 2024 Sep 6;15(1):7722. doi: 10.1038/s41467-024-52107-w. PMID:39242571[33]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

  1. ↑ Dogra S, Krishnamurthy S, Gupta V, Dixit BL, Gupta CM, Sanglard D, Prasad R. Asymmetric distribution of phosphatidylethanolamine in C. albicans: possible mediation by CDR1, a multidrug transporter belonging to ATP binding cassette (ABC) superfamily. Yeast. 1999 Jan 30;15(2):111-21. PMID:10029989 doi:<111::AID-YEA350>3.0.CO;2-E 10.1002/(SICI)1097-0061(19990130)15:2<111::AID-YEA350>3.0.CO;2-E
  2. ↑ Smriti, Krishnamurthy SS, Prasad R. Membrane fluidity affects functions of Cdr1p, a multidrug ABC transporter of Candida albicans. FEMS Microbiol Lett. 1999 Apr 15;173(2):475-81. PMID:10227177 doi:10.1111/j.1574-6968.1999.tb13541.x
  3. ↑ Henry KW, Cruz MC, Katiyar SK, Edlind TD. Antagonism of azole activity against Candida albicans following induction of multidrug resistance genes by selected antimicrobial agents. Antimicrob Agents Chemother. 1999 Aug;43(8):1968-74. PMID:10428921 doi:10.1128/AAC.43.8.1968
  4. ↑ Nakamura K, Niimi M, Niimi K, Holmes AR, Yates JE, Decottignies A, Monk BC, Goffeau A, Cannon RD. Functional expression of Candida albicans drug efflux pump Cdr1p in a Saccharomyces cerevisiae strain deficient in membrane transporters. Antimicrob Agents Chemother. 2001 Dec;45(12):3366-74. PMID:11709310 doi:10.1128/AAC.45.12.3366-3374.2001
  5. ↑ Smriti, Krishnamurthy S, Dixit BL, Gupta CM, Milewski S, Prasad R. ABC transporters Cdr1p, Cdr2p and Cdr3p of a human pathogen Candida albicans are general phospholipid translocators. Yeast. 2002 Mar 15;19(4):303-18. PMID:11870854 doi:10.1002/yea.818
  6. ↑ Gauthier C, Weber S, Alarco AM, Alqawi O, Daoud R, Georges E, Raymond M. Functional similarities and differences between Candida albicans Cdr1p and Cdr2p transporters. Antimicrob Agents Chemother. 2003 May;47(5):1543-54. PMID:12709320 doi:10.1128/AAC.47.5.1543-1554.2003
  7. ↑ Shukla S, Saini P, Smriti, Jha S, Ambudkar SV, Prasad R. Functional characterization of Candida albicans ABC transporter Cdr1p. Eukaryot Cell. 2003 Dec;2(6):1361-75. PMID:14665469 doi:10.1128/EC.2.6.1361-1375.2003
  8. ↑ O'Keeffe J, Kavanagh K. Adriamycin alters the expression of drug efflux pumps and confers amphotericin B tolerance in Candida albicans. Anticancer Res. 2004 Mar-Apr;24(2A):405-8 PMID:15152937
  9. ↑ Shukla S, Ambudkar SV, Prasad R. Substitution of threonine-1351 in the multidrug transporter Cdr1p of Candida albicans results in hypersusceptibility to antifungal agents and threonine-1351 is essential for synergic effects of calcineurin inhibitor FK520. J Antimicrob Chemother. 2004 Jul;54(1):38-45. PMID:15190023 doi:10.1093/jac/dkh308
  10. ↑ Niimi M, Niimi K, Takano Y, Holmes AR, Fischer FJ, Uehara Y, Cannon RD. Regulated overexpression of CDR1 in Candida albicans confers multidrug resistance. J Antimicrob Chemother. 2004 Dec;54(6):999-1006. PMID:15486081 doi:10.1093/jac/dkh456
  11. ↑ Saini P, Prasad T, Gaur NA, Shukla S, Jha S, Komath SS, Khan LA, Haq QM, Prasad R. Alanine scanning of transmembrane helix 11 of Cdr1p ABC antifungal efflux pump of Candida albicans: identification of amino acid residues critical for drug efflux. J Antimicrob Chemother. 2005 Jul;56(1):77-86. PMID:15937063 doi:10.1093/jac/dki183
  12. ↑ Tanida T, Okamoto T, Ueta E, Yamamoto T, Osaki T. Antimicrobial peptides enhance the candidacidal activity of antifungal drugs by promoting the efflux of ATP from Candida cells. J Antimicrob Chemother. 2006 Jan;57(1):94-103. PMID:16291868 doi:10.1093/jac/dki402
  13. ↑ Shukla S, Rai V, Banerjee D, Prasad R. Characterization of Cdr1p, a major multidrug efflux protein of Candida albicans: purified protein is amenable to intrinsic fluorescence analysis. Biochemistry. 2006 Feb 21;45(7):2425-35. PMID:16475832 doi:10.1021/bi0519147
  14. ↑ Saini P, Gaur NA, Prasad R. Chimeras of the ABC drug transporter Cdr1p reveal functional indispensability of transmembrane domains and nucleotide-binding domains, but transmembrane segment 12 is replaceable with the corresponding homologous region of the non-drug transporter Cdr3p. Microbiology (Reading). 2006 May;152(Pt 5):1559-1573. PMID:16622073 doi:10.1099/mic.0.28471-0
  15. ↑ Gao PH, Cao YB, Jia XM, Cao YY, Quan H, Wang Y, Jiang YY. Drug susceptibilities of yeast cells are affected when expressing mutant Candida albicans drug resistance protein. Int J Antimicrob Agents. 2006 Jul;28(1):69-74. PMID:16782311 doi:10.1016/j.ijantimicag.2006.02.016
  16. ↑ Shen H, An MM, Wang de J, Xu Z, Zhang JD, Gao PH, Cao YY, Cao YB, Jiang YY. Fcr1p inhibits development of fluconazole resistance in Candida albicans by abolishing CDR1 induction. Biol Pharm Bull. 2007 Jan;30(1):68-73. PMID:17202662 doi:10.1248/bpb.30.68
  17. ↑ Pasrija R, Panwar SL, Prasad R. Multidrug transporters CaCdr1p and CaMdr1p of Candida albicans display different lipid specificities: both ergosterol and sphingolipids are essential for targeting of CaCdr1p to membrane rafts. Antimicrob Agents Chemother. 2008 Feb;52(2):694-704. PMID:18056285 doi:10.1128/AAC.00861-07
  18. ↑ Manoharlal R, Gaur NA, Panwar SL, Morschhäuser J, Prasad R. Transcriptional activation and increased mRNA stability contribute to overexpression of CDR1 in azole-resistant Candida albicans. Antimicrob Agents Chemother. 2008 Apr;52(4):1481-92. PMID:18268086 doi:10.1128/AAC.01106-07
  19. ↑ Holmes AR, Lin YH, Niimi K, Lamping E, Keniya M, Niimi M, Tanabe K, Monk BC, Cannon RD. ABC transporter Cdr1p contributes more than Cdr2p does to fluconazole efflux in fluconazole-resistant Candida albicans clinical isolates. Antimicrob Agents Chemother. 2008 Nov;52(11):3851-62. PMID:18710914 doi:10.1128/AAC.00463-08
  20. ↑ Tsao S, Rahkhoodaee F, Raymond M. Relative contributions of the Candida albicans ABC transporters Cdr1p and Cdr2p to clinical azole resistance. Antimicrob Agents Chemother. 2009 Apr;53(4):1344-52. PMID:19223631 doi:10.1128/AAC.00926-08
  21. ↑ Puri N, Gaur M, Sharma M, Shukla S, Ambudkar SV, Prasad R. The amino acid residues of transmembrane helix 5 of multidrug resistance protein CaCdr1p of Candida albicans are involved in substrate specificity and drug transport. Biochim Biophys Acta. 2009 Sep;1788(9):1752-61. PMID:19393219 doi:10.1016/j.bbamem.2009.04.009
  22. ↑ Sun LM, Cheng AX, Wu XZ, Zhang HJ, Lou HX. Synergistic mechanisms of retigeric acid B and azoles against Candida albicans. J Appl Microbiol. 2010 Jan;108(1):341-8. PMID:20002912 doi:10.1111/j.1365-2672.2009.04429.x
  23. ↑ Basso LR Jr, Gast CE, Mao Y, Wong B. Fluconazole transport into Candida albicans secretory vesicles by the membrane proteins Cdr1p, Cdr2p, and Mdr1p. Eukaryot Cell. 2010 Jun;9(6):960-70. PMID:20348384 doi:10.1128/EC.00355-09
  24. ↑ Siikala E, Rautemaa R, Richardson M, Saxen H, Bowyer P, Sanglard D. Persistent Candida albicans colonization and molecular mechanisms of azole resistance in autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) patients. J Antimicrob Chemother. 2010 Dec;65(12):2505-13. PMID:20876623 doi:10.1093/jac/dkq354
  25. ↑ Prasad R, Shah AH, Sanwal H, Kapoor K. Alanine scanning of all cysteines and construction of a functional cysteine-less Cdr1p, a multidrug ABC transporter of Candida albicans. Biochem Biophys Res Commun. 2012 Jan 6;417(1):508-13. PMID:22166216 doi:10.1016/j.bbrc.2011.11.150
  26. ↑ Zhu J, Krom BP, Sanglard D, Intapa C, Dawson CC, Peters BM, Shirtliff ME, Jabra-Rizk MA. Farnesol-induced apoptosis in Candida albicans is mediated by Cdr1-p extrusion and depletion of intracellular glutathione. PLoS One. 2011;6(12):e28830. PMID:22205973 doi:10.1371/journal.pone.0028830
  27. ↑ Rawal MK, Khan MF, Kapoor K, Goyal N, Sen S, Saxena AK, Lynn AM, Tyndall JD, Monk BC, Cannon RD, Komath SS, Prasad R. Insight into pleiotropic drug resistance ATP-binding cassette pump drug transport through mutagenesis of Cdr1p transmembrane domains. J Biol Chem. 2013 Aug 23;288(34):24480-93. PMID:23824183 doi:10.1074/jbc.M113.488353
  28. ↑ Thomas E, Roman E, Claypool S, Manzoor N, Pla J, Panwar SL. Mitochondria influence CDR1 efflux pump activity, Hog1-mediated oxidative stress pathway, iron homeostasis, and ergosterol levels in Candida albicans. Antimicrob Agents Chemother. 2013 Nov;57(11):5580-99. PMID:23979757 doi:10.1128/AAC.00889-13
  29. ↑ Sanglard D, Ischer F, Monod M, Bille J. Susceptibilities of Candida albicans multidrug transporter mutants to various antifungal agents and other metabolic inhibitors. Antimicrob Agents Chemother. 1996 Oct;40(10):2300-5. PMID:8891134 doi:10.1128/AAC.40.10.2300
  30. ↑ Albertson GD, Niimi M, Cannon RD, Jenkinson HF. Multiple efflux mechanisms are involved in Candida albicans fluconazole resistance. Antimicrob Agents Chemother. 1996 Dec;40(12):2835-41. PMID:9124851 doi:10.1128/AAC.40.12.2835
  31. ↑ Krishnamurthy S, Gupta V, Snehlata P, Prasad R. Characterisation of human steroid hormone transport mediated by Cdr1p, a multidrug transporter of Candida albicans, belonging to the ATP binding cassette super family. FEMS Microbiol Lett. 1998 Jan 1;158(1):69-74. PMID:9453158 doi:10.1111/j.1574-6968.1998.tb12802.x
  32. ↑ Krishnamurthy S, Chatterjee U, Gupta V, Prasad R, Das P, Snehlata P, Hasnain SE, Prasad R. Deletion of transmembrane domain 12 of CDR1, a multidrug transporter from Candida albicans, leads to altered drug specificity: expression of a yeast multidrug transporter in baculovirus expression system. Yeast. 1998 Apr 30;14(6):535-50. PMID:9605504 doi:<535::AID-YEA254>3.0.CO;2-5 10.1002/(SICI)1097-0061(19980430)14:6<535::AID-YEA254>3.0.CO;2-5
  33. ↑ Peng Y, Lu Y, Sun H, Ma J, Li X, Han X, Fang Z, Tan J, Qiu Y, Qu T, Yin M, Yan Z. Cryo-EM structures of Candida albicans Cdr1 reveal azole-substrate recognition and inhibitor blocking mechanisms. Nat Commun. 2024 Sep 6;15(1):7722. PMID:39242571 doi:10.1038/s41467-024-52107-w

9ium, resolution 3.08Å

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