Short Communication

Molecular Characterization of Aquaglyceroporine: A Novel Mutation in LmAQP1 from Leishmania major (MRHO/IR/75/ER)


Background: The first line treatment for cutaneous leishmaniasis is pentavalent antimony such as sodium stibogluconate (pentostam) and meglumine antimonite (glucantime). One of the most important ways to uptake the drug is by a transmembrane protein, called aquaglyceroporin encoded by Aquaglyceroprotein1 (LmAQP1). In this study, molecular characterization of LmAQP1 was reported.

Methods: Leishmania major (MRHO/IR/75/ER) promastigotes were cultured, and then DNA extraction and RNA extraction were done and followed by cDNA synthesis. Amplicons resulted from PCR and RT-PCR using specific primers were purified and sequenced. Molecular characterization was done by bioinformatically software such as BLST, ClustalW2, and RMSD.

Results: Amplicons resulted from PCR and RT-PCR showed equal size in length. BLASTn analysis showed a point nucleotide change in LmAQP1 gene that encoded 282-amino-acid long protein with a mutation at position 154 including replacement of alanine by threonine. The observed mutation in the interested gene was assessed using the above-mentioned software. The mentioned gene was submitted at GenBank, NCBI with accession number of KU514052.

Conclusion: The functional prediction of the protein encoded from LmAQP1 showed that the mentioned mutation could not affect the three-dimension structure, but it may modify the drug uptake potential of this important channel. Based on from LmAQP1 role, it seems to be an appropriate candidate for drug development. According to search through internet, this is the first report of LmAQP1 from L. major (MRHO/IR/75/ER).

1. Desjeux P. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis. 2004;27(5):305-18.
2. Kedzierski L, Sakthianandeswaren A, Curtis JM et al. Leishmaniasis: current treatment and prospects for new drugs and vaccines. Curr Med Chem. 2009;16(5):599-614.
3. Gourbal B, Sonuc N, Bhattacharjee H et al. Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin. J Biol Chem. 2004;279(30):31010-7.
4. Kozono D, Yasui M, King LS, Agre P. Aquaporin water channels: atomic structure molecular dynamics meet clinical medicine. J Clin Invest. 2002;109(11):1395-9.
5. Marquis N, Gourbal B, Rosen BP et al. Modulation in aquaglyceroporin AQP1 gene transcript levels in drug‐resistant Leishmania. Mol Microbiol. 2005;57(6):1690-9.
6. Figarella K, Uzcategui NL, Zhou Y et al. Biochemical characterization of Leishmania major aquaglyceroporin LmAQP1: possible role in volume regulation and osmotaxis. Mol Microbiol. 2007;65(4):1006-17.
7. Croft SL, Sundar S, Fairlamb AH. Drug resistance in leishmaniasis. Clin Microbiol Rev. 2006;19(1):111-26.
8. Gómez Pérez V, García-Hernandez R, Corpas-López V et al. Decreased antimony uptake and overexpression of genes of thiol metabolism are associated with drug resistance in a canine isolate of Leishmania infantum. Int J Parasitol Drugs Drug Resist. 2016;6(2):133-9.
9. Mandal G, Sarkar A, Saha P, Singh N et al. Func-tionality of drug efflux pumps in antimonial re-sistant Leishmania donovani field isolates. Indian J Bio-chem Biophys. 2009;46(1):86-92.
10. Beverley SM. Gene amplification in Leishmania. Annu Rev Microbiol. 1991;45:417-44.
11. Ouellette M, Hettema E, Wüst D, Fase-Fowler F, Borst P. Direct and inverted DNA repeats associated with P-glycoprotein gene amplification in drug resistant Leishmania. EMBO J. 1991;10(4):1009-16.
12. Hadighi R, Mohebali M, Boucher P et al. Unre-spon-siveness to Glucantime treatment in Iranian cutaneous leishmaniasis due to drug resistant Leish-mania tropica parasites. PLoS Med. 2006;3(5):e162.
13. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB et al. Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol. 2013;135(2):344-9.
14. Zarean M, Maraghi S, Hajjaran H et al. Compari-son of proteome profiling of two sensitive and re-sistant field Iranian isolates of Leishmania major to Glucantime® by 2- Dimensional electrophoresis. Iran J Parasitol. 2015;10(1):19-29.
15. Khamesipour A, Dowlati Y, Asilian A et al. Leishmanization: use of an old method for evaluation of candidate vaccines against leishmaniasis. Vaccine. 2005;23(28):3642-8.
16. Eisenberger CL, Jaffe CL. Leishmania: identification of Old World species using a permissively primed intergenic polymorphic-polymerase chain reaction. Exp Parasitol. 1999;91(1):70-7.
17. Altschul SF, Madden TL, Schäffer AA et al. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997;25(17):3389-402.
18. Altschul SF, Wootton JC, Gertz EM et al. Protein database searches using compositionally adjusted substitution matrices. FEBS J. 2005;272(20):5101-9.
19. Eslami G, Salehi R. Genetic variation in RPOIILS gene encoding RNA polymerase II largest subunit from Leishmania major. Mol Biol Rep. 2014;41(4):2585-9.
20. Eslami G, Salehi R, Khosravi S, Doudi M. Genetic analysis of clinical isolates of Leishmania major from Isfahan, Iran. J Vector Borne Dis. 2012;49(3):168-74.
21. Eslami G, Anvarii H, Ebadi M et al. The changing profile of cutaneous leishmaniasis agent in a central province of Iran. Tanzan J Health Res. 2013;15(1):33-9.
22. Mukhopadhyay R. An Aquaglyceroporin as a New Drug Target in Leishmania. Clin Res Infect Dis. 2013; 1(1): 1001.
23. Murata K, Mitsuoka K, Hirai T et al. Structural de-terminants of water permeation through aqua-porin-1. Nature. 2000;407(6804):599-605.
24. Mukhopadhyay R, Mandal G, Atluri VS et al. The role of alanine 163 in solute permeability of Leishma-nia major aquaglyceroporin LmAQP1. Mol Bio-chem Parasitol. 2011;175(1):83-90.
25. Zee RY, Cook NR, Cheng S et al. Threonine for alanine substitution in the eotaxin (CCL11) gene and the risk of incident myocardial infarction. Atherosclerosis. 2004;175(1):91-4.
26. Podoly E, Hanin G, Soreq H. Alanine-to-threonine substitutions and amyloid diseases: Butyrylcholinesterase as a case study. Chem Biol Interact. 2010;187(1-3):64-71.
27. Monte-Neto R, Laffitte MC, Leprohon P et al. Intrachromosomal amplification, locus deletion and point mutation in the aquaglyceroporin AQP1 gene in antimony resistant Leishmania (Viannia) guya-nensis. PLoS Negl Trop Dis. 2015;9(2):e0003476.
IssueVol 14 No 3 (2019) QRcode
SectionShort Communication(s)
Aquaporin 1 Leishmania Molecular dynamics simulation Antimony

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
ESLAMI G, GHAVAMI M, MORADI AR, NADRI H, AHMADIAN S. Molecular Characterization of Aquaglyceroporine: A Novel Mutation in LmAQP1 from Leishmania major (MRHO/IR/75/ER). Iran J Parasitol. 2019;14(3):465-471.