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Detection of Mycolactone by Thin Layer Chromatography

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Mycobacterium ulcerans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2387))

Abstract

By means of thin layer chromatography coupled to a fluorescence enhancer, a highly sensitive and operationally simple method to detect the mycolactones stemming from the human pathogen Mycobacterium ulcerans was developed and applied to various sample sources.

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References

  1. Chany AC, Tresse C, Casarotto V, Blanchard N (2013) History, biology and chemistry of mycobacterium ulcerans infections (Buruli ulcer disease). Nat Prod Rep 30:1527–1567. https://doi.org/10.1039/c3np70068b

    Article  CAS  PubMed  Google Scholar 

  2. Adusumilli S, Mve-Obiang A, Sparer T, Meyers W, Hayman J, Small PL (2005) Mycobacterium ulcerans toxic macrolide, mycolactone modulates the host immune response and cellular location of M. ulcerans in vitro and in vivo. Cell Microbiol 7:1295–1304

    Article  CAS  Google Scholar 

  3. Snyder DS, Small PL (2003) Uptake and cellular actions of mycolactone, a virulence determinant for mycobacterium ulcerans. Microb Pathog 34:91–101

    Article  CAS  Google Scholar 

  4. Ranger BS, Mahrous EA, Mosi L, Adusumilli S, Lee RE, Colorni A, Rhodes M, Small PL (2006) Globally distributed mycobacterial fish pathogens produce a novel plasmid-encoded toxic macrolide, mycolactone F. Infect Immun 74:6037–6045

    Article  CAS  Google Scholar 

  5. Sarfo FS, Phillips RO, Rangers B, Mahrous EA, Lee RE, Tarelli E, Asiedu KB, Small PL, Wansbrough-Jones MH (2010) Detection of Mycolactone A/B in mycobacterium ulcerans-infected human tissue. PLoS Negl Trop Dis 4(1):e577. https://doi.org/10.1371/journal.pntd.0000577

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Song F, Fidanze S, Benowitz AB, Kishi Y (2007) Total synthesis of Mycolactones A and B. Tetrahedron 6:5739–5753

    Article  Google Scholar 

  7. George KM, Chatterjee D, Gunawardana G, Welty D, Hayman J, Lee R, Small PL (1999) Mycolactone: a polyketide toxin from Mycobacterium ulcerans required for virulence. Science 283:854–857

    Article  CAS  Google Scholar 

  8. George KM, Pascopella L, Welty DM, Small PL (2000) A Mycobacterium ulcerans toxin, mycolactone, causes apoptosis in Guinea pig ulcers and tissue culture cells. Infect Immun 68:877–883

    Article  CAS  Google Scholar 

  9. Staunton J, Weissman KJ (2001) Polyketide biosynthesis: a millennium review. Nat Prod Rep 18:380–416

    Article  CAS  Google Scholar 

  10. Pidot SJ, Hong H, Seemann T, Porter JL, Yip MJ, Men A, Johnson M, Wilson P, Davies JK, Leadlay PF, Stinear TP (2008) Deciphering the genetic basis for polyketide variation among mycobacteria producing mycolactones. BMC Genomics 9:462. https://doi.org/10.1186/1471-2164-9-462

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hopwood DA (1997) Genetic contributions to understanding polyketide synthases. Chem Rev 97:2465–2498

    Article  CAS  Google Scholar 

  12. Khosla C (2009) Structures and mechanisms of polyketide synthases. J Org Chem 74:6416–6420. https://doi.org/10.1021/jo9012089

    Article  CAS  PubMed  Google Scholar 

  13. Hong H, Stinear T, Porter J, Demangel C, Leadlay PF (2007) A novel mycolactone toxin obtained by biosynthetic engineering. Chembiochem 8:2043–2047

    Article  CAS  Google Scholar 

  14. Mve-Obiang A, Lee RE, Portaels F, Small PLC (2003) Heterogeneity of Mycolactones produced by clinical isolates of mycobacterium ulcerans: implications for virulence. Infect Immun 71:774–783

    Article  CAS  Google Scholar 

  15. Ranger BS, Mahrous EA, Mosi L, Adusumilli S, Lee RE, Colorni A, Rhodes M, Small PLC (2006) Globally distributed mycobacterial fish pathogens produce a novel plasmid-encoded toxic macrolide. Mycolactone F Infect Immun 74:6037–6045

    Article  CAS  Google Scholar 

  16. Gunawardana G, Chatterjee D, George KM, Brennan P, Whittern D, Small PLC (1999) Characterization of novel macrolide toxins, Mycolactones a and B, from a human pathogen, mycobacterium ulcerans. J Am Chem Soc 121:6092–6093

    Article  CAS  Google Scholar 

  17. Benowitz AB, Fidanze S, Small PL, Kishi Y (2001) Stereochemistry of the core structure of the mycolactones. J Am Chem Soc 123:5128–5129

    Article  CAS  Google Scholar 

  18. Fidanze S, Song F, Szlosek-Pinaud M, Small PL, Kishi Y (2001) Complete structure of the mycolactones. J Am Chem Soc 123:10117–10118

    Article  CAS  Google Scholar 

  19. Higashibayashi S, Czechtizky W, Kobayashi Y, Kishi Y (2003) Universal NMR databases for contiguous polyols. J Am Chem Soc 125:14379–14393

    Article  CAS  Google Scholar 

  20. Seike H, Ghosh I, Kishi Y (2006) Attempts to assemble a universal NMR database without synthesis of NMR database compounds. Org Lett 8:3861–3864

    Article  CAS  Google Scholar 

  21. Fleury E, Lannou MI, Bistri O, Sautel F, Massiot G, Pancrazi A, Ardisson J (2009) Relative stereochemical determination and synthesis of the C1-C17 fragment of a new natural polyketide. J Org Chem 74:7034–7045. https://doi.org/10.1021/jo9012833

    Article  CAS  PubMed  Google Scholar 

  22. Thin-Layer Chromatography—TLC Introduction https://www.merckmillipore.com/DE/en/analytics-sample-preparation/learning-center-thin-layer-chromatography/59Ob.qB.emsAAAFVa.5Dx06W,nav. Accessed Feb 24 2020

  23. Maximov OB, Panthinkhina LS (1965) Potassium permanganate alkaline for reducing compounds and aromatic polycarboxylic acids. Spray solution: Add to 1% aqueous potassium permanganate solution an equal volume of 5% aqueous sodium carbonate solution. J Chromatogr A 150:20

    Google Scholar 

  24. Stahl E, Kaltenbach U (1961) Anisaldehyde—sulfuric acid for sugars, steroids, terpenes. Spray solution: Prepare freshly before use a solution of 0.5 mL anisaldehyde in 50 mL glacial acetic acid and 1 mL 97% sulfuric acid. After-treatment: Heat to 100-105 °C until maximal visualisation of the spots. The background may be brightened by water vapour. J Chromatogr A 5:351

    Article  CAS  Google Scholar 

  25. Kuivila GH, Keough AH, Soboczenski EJ (1954) Areneboronates from diols and polyols. J Org Chem 19:780–783

    Google Scholar 

  26. Trop M, Sprecher M, Pinsky A (1968) Ammonium cerium(IV) nitrate—nitric acid for α-hydroxy acids, α-keto acids and mercaptans. Dip solution: Dissolve 20 g ammonium cerium(IV) nitrate in 50 mL 0.5 N nitric acid. Dilute freshly before use 1 part of this solution with 3 parts water. Procedure: After drying dip the chromatogram into the dip solution and place it on a clean filter paper. White spots on yellow background. J Chromatogr A 32:426

    Article  CAS  Google Scholar 

  27. de Silva AP, Gunaratne HQ, Gunnlaugsson T, Huxley AJ, McCoy CP, Rademacher JT, Rice TE (1997) Signaling recognition events with fluorescent sensors and switches. Chem Rev 97:1515–1566

    Article  Google Scholar 

  28. Kuivila GH, Keough AH, Soboczenski EJ (1954) Areneboronates from diols and polyols. J Org Chem 19:780–783

    Article  CAS  Google Scholar 

  29. James TD, Shinkai S (2002) Artificial receptors as Chemosensors for carbohydrates. In: Penadés S (ed) Host-guest chemistry. Topics in current chemistry, vol 218. Springer, Berlin, Heidelberg

    Google Scholar 

  30. Suenaga H, Mikami M, Sandanayake S, Shinkai S (1995) Screening of fluorescent boronic acids for sugar sensing which show a large fluorescence change. Tetrahedron Lett 36:4825

    Article  CAS  Google Scholar 

  31. Springsteen G, Wang B (2002) A detailed examination of boronic acid–diol complexation. Tetrahedron 58:5291–5300

    Article  CAS  Google Scholar 

  32. Spangenberg T, Kishi Y (2010) Highly sensitive, operationally simple, cost/time effective detection of the mycolactones from the human pathogen mycobacterium ulcerans. Chem Commun 46:1410–1412

    Article  CAS  Google Scholar 

  33. Converse PJ, Xing Y, Kim KH, Tyagi S, Li SY, Almeida DV, Nuermberger EL, Grosset JH, Kishi Y (2014) Accelerated detection of mycolactone production and response to antibiotic treatment in a mouse model of mycobacterium ulcerans disease. PLoS Negl Trop Dis 8:e2618. https://doi.org/10.1371/journal.pntd.0002618

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Wadagni A, Frimpong M, Phanzu DM, Ablordey A, Kacou E, Gbedevi M et al (2015) Simple, rapid Mycobacterium ulcerans disease diagnosis from clinical samples by fluorescence of mycolactone on thin layer chromatography. PLoS Negl Trop Dis 9(11):e0004247

    Article  Google Scholar 

  35. https://www.youtube.com/watch?v=6RoHYCYO8rwl. Accessed Feb 24 2020

  36. https://www.youtube.com/watch?v=8SF4jNe3aSc. Accessed Feb 24 2020

  37. Folch J, Lees M, Sloane SGH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226:497–509

    Article  CAS  Google Scholar 

  38. Sarfo FS, Phillips RO, Zhang J, Abass MK, Abotsi J, Amoako YA, Adu-Sarkodie Y, Robinson C, Wansbrough-Jones MH (2014) Kinetics of mycolactone in human subcutaneous tissue during antibiotic therapy for mycobacterium ulcerans disease. BMC Infect Dis 14:202. https://doi.org/10.1186/1471-2334-14-202

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Pettit JHS, Marchette NJ, Rees RJW (1966) Mycobacterium ulcerans infection. Clinical and bacteriological study of the first cases recognized in South East Asia. Br J Dermatol 78:187–197

    Article  CAS  Google Scholar 

  40. Dega H, Bentoucha A, Robert J, Jarlier V, Grosset J (2002) Bactericidal activity of rifampin-amikacin against mycobacterium ulcerans in mice. Antimicrob Agents Chemother 46:3193–3196

    Article  CAS  Google Scholar 

  41. Takemura T, Chihara K, Becker RS, Das PK, Hug GL (1980) Visual pigments. 11. Spectroscopy and photophysics of retinoic acids and all-trans-methyl retinoate. J Am Chem Soc 102:2604–2609

    Article  CAS  Google Scholar 

  42. Marion E, Prado S, Cano C, Babonneau J, Ghamrawi S, Marsollier L (2012) Photodegradation of the mycobacterium ulcerans toxin, Mycolactones: considerations for handling and storage. PLoS One 7:e33600. https://doi.org/10.1371/journal.pone.0033600

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Xing Y, Hande SM, Kishi Y (2012) Photochemistry of Mycolactone A/B, the causative toxin of Buruli ulcer. J Am Chem Soc 134:19234–19239

    Article  CAS  Google Scholar 

  44. Lamola AA, Leermakers PA, Byers GW, Hammond GS (1965) Intramolecular electronic energy transfer between nonconjugated chromophores in some model compounds. J Am Chem Soc 87:2322–2332

    Article  CAS  Google Scholar 

  45. Shiratori H, Ohno T, Nozaki K, Osuka A (2000) Coordination control of intramolecular energy transfer in boronate-bridged naphthalene-aryl ketone molecule. Chem Phys Lett 320:631–638

    Article  CAS  Google Scholar 

  46. Kubicek-Sutherland JZ, Vu DM, Anderson AS, Mukundan H, Sanchez TC, Marti-Arbona R, Swanson BI, Converse PJ, Nuermberger EL (2019) Understanding the significance of biochemistry in the storage, handling, purification, and sampling of amphiphilic Mycolactone. Toxins 11:202

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Chemistry, University of Ghana, Legon, Ghana

    Richard Kwamla Amewu

  2. Global Health Institute of Merck, Ares Trading S.A. (a subsidiary of Merck KGaA Darmstadt Germany), Eysins, Switzerland

    Thomas Spangenberg

Authors
  1. Richard Kwamla Amewu
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  2. Thomas Spangenberg
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Corresponding author

Correspondence to Richard Kwamla Amewu .

Editor information

Editors and Affiliations

  1. Swiss Tropical and Public Health Institute, Basel, Switzerland

    Gerd Pluschke

  2. Stanford School of Medicine Department of Pathology, Stanford University, Stanford, CA, USA

    Katharina Röltgen

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Amewu, R.K., Spangenberg, T. (2022). Detection of Mycolactone by Thin Layer Chromatography. In: Pluschke, G., Röltgen, K. (eds) Mycobacterium ulcerans. Methods in Molecular Biology, vol 2387. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1779-3_14

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  • DOI: https://doi.org/10.1007/978-1-0716-1779-3_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1778-6

  • Online ISBN: 978-1-0716-1779-3

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