Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone
Abstract
Mycolactone, the amphiphilic macrolide toxin secreted by Mycobacterium ulcerans, plays a significant role in the pathology and manifestations of Buruli ulcer (BU). Consequently, it follows that the toxin is a suitable target for the development of diagnostics and therapeutics for this disease. Yet, several challenges have deterred such development. For one, the lipophilic nature of the toxin makes it difficult to handle and store and contributes to variability associated with laboratory experimentation and purification yields. In this manuscript, we have attempted to incorporate our understanding of the lipophilicity of mycolactone in order to define the optimal methods for the storage, handling, and purification of this toxin. We present a systematic correlation of variability associated with measurement techniques (thin-layer chromatography (TLC), mass spectrometry (MS), and UV-Vis spectrometry), storage conditions, choice of solvents, as well as the impact of each of these on toxin function as assessed by cellular cytotoxicity. We also compared natural mycolactone extracted from bacterial culture with synthesized toxins in laboratory (solvents, buffers) and physiologically relevant (serum) matrices. Our results point to the greater stability of mycolactone in organic, as well as detergent-containing, solvents, regardless of the container material (plastic, glass, or silanized tubes). They also highlight the presencemore »
- Authors:
-
- Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
- Johns Hopkins Univ. Center for Tuberculosis Research, Baltimore, MD (United States)
- Publication Date:
- Research Org.:
- Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
- Sponsoring Org.:
- National Institutes of Health (NIH); USDOE
- OSTI Identifier:
- 1511262
- Report Number(s):
- LA-UR-19-20386
Journal ID: ISSN 2072-6651; TOXIB7
- Grant/Contract Number:
- 89233218CNA000001
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Toxins
- Additional Journal Information:
- Journal Volume: 11; Journal Issue: 4; Journal ID: ISSN 2072-6651
- Publisher:
- MDPI
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Biological Science; mycolactone; storage and handling; amphiphilic; lipoproteins
Citation Formats
Kubicek-Sutherland, Jessica Zofie, Vu, Dung M., Anderson, Aaron S., Sanchez, Timothy Charles, Converse, Paul J., Martí-Arbona, Ricardo, Nuermberger, Eric L., Swanson, Basil Ian, and Mukundan, Harshini. Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone. United States: N. p., 2019.
Web. doi:10.3390/toxins11040202.
Kubicek-Sutherland, Jessica Zofie, Vu, Dung M., Anderson, Aaron S., Sanchez, Timothy Charles, Converse, Paul J., Martí-Arbona, Ricardo, Nuermberger, Eric L., Swanson, Basil Ian, & Mukundan, Harshini. Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone. United States. https://doi.org/10.3390/toxins11040202
Kubicek-Sutherland, Jessica Zofie, Vu, Dung M., Anderson, Aaron S., Sanchez, Timothy Charles, Converse, Paul J., Martí-Arbona, Ricardo, Nuermberger, Eric L., Swanson, Basil Ian, and Mukundan, Harshini. Thu .
"Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone". United States. https://doi.org/10.3390/toxins11040202. https://www.osti.gov/servlets/purl/1511262.
@article{osti_1511262,
title = {Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone},
author = {Kubicek-Sutherland, Jessica Zofie and Vu, Dung M. and Anderson, Aaron S. and Sanchez, Timothy Charles and Converse, Paul J. and Martí-Arbona, Ricardo and Nuermberger, Eric L. and Swanson, Basil Ian and Mukundan, Harshini},
abstractNote = {Mycolactone, the amphiphilic macrolide toxin secreted by Mycobacterium ulcerans, plays a significant role in the pathology and manifestations of Buruli ulcer (BU). Consequently, it follows that the toxin is a suitable target for the development of diagnostics and therapeutics for this disease. Yet, several challenges have deterred such development. For one, the lipophilic nature of the toxin makes it difficult to handle and store and contributes to variability associated with laboratory experimentation and purification yields. In this manuscript, we have attempted to incorporate our understanding of the lipophilicity of mycolactone in order to define the optimal methods for the storage, handling, and purification of this toxin. We present a systematic correlation of variability associated with measurement techniques (thin-layer chromatography (TLC), mass spectrometry (MS), and UV-Vis spectrometry), storage conditions, choice of solvents, as well as the impact of each of these on toxin function as assessed by cellular cytotoxicity. We also compared natural mycolactone extracted from bacterial culture with synthesized toxins in laboratory (solvents, buffers) and physiologically relevant (serum) matrices. Our results point to the greater stability of mycolactone in organic, as well as detergent-containing, solvents, regardless of the container material (plastic, glass, or silanized tubes). They also highlight the presence of toxin in samples that may be undetectable by any one technique, suggesting that each detection approach captures different configurations of the molecule with varying specificity and sensitivity. Most importantly, our results demonstrate for the very first time that amphiphilic mycolactone associates with host lipoproteins in serum, and that this association will likely impact our ability to study, diagnose, and treat Buruli ulcers in patients.},
doi = {10.3390/toxins11040202},
journal = {Toxins},
number = 4,
volume = 11,
place = {United States},
year = {2019},
month = {4}
}
Web of Science
Figures / Tables:

Works referenced in this record:
Accelerated Detection of Mycolactone Production and Response to Antibiotic Treatment in a Mouse Model of Mycobacterium ulcerans Disease
journal, January 2014
- Converse, Paul J.; Xing, Yalan; Kim, Ki Hyun
- PLoS Neglected Tropical Diseases, Vol. 8, Issue 1
Total Synthesis of the Mycolactones
journal, February 2002
- Song, Fengbin; Fidanze, Steve; Benowitz, Andrew B.
- Organic Letters, Vol. 4, Issue 4
The potent effect of mycolactone on lipid membranes
journal, January 2018
- Nitenberg, Milène; Bénarouche, Anaïs; Maniti, Ofelia
- PLOS Pathogens, Vol. 14, Issue 1
Immunosuppressive Properties of the Soluble Toxin from Mycobacterium ulcerans
journal, March 1988
- Pimsler, M.; Sponsler, T. A.; Meyers, W. M.
- Journal of Infectious Diseases, Vol. 157, Issue 3
The chemistry and biology of mycolactones
journal, January 2017
- Gehringer, Matthias; Altmann, Karl-Heinz
- Beilstein Journal of Organic Chemistry, Vol. 13
A Ring-Closing Metathesis (RCM)-Based Approach to Mycolactones A/B
journal, October 2011
- Gersbach, Philipp; Jantsch, Andrea; Feyen, Fabian
- Chemistry - A European Journal, Vol. 17, Issue 46
Complete Structure of the Mycolactones
journal, October 2001
- Fidanze, Steve; Song, Fengbin; Szlosek-Pinaud, Magali
- Journal of the American Chemical Society, Vol. 123, Issue 41
Simple, Rapid Mycobacterium ulcerans Disease Diagnosis from Clinical Samples by Fluorescence of Mycolactone on Thin Layer Chromatography
journal, November 2015
- Wadagni, Anita; Frimpong, Michael; Phanzu, Delphin Mavinga
- PLOS Neglected Tropical Diseases, Vol. 9, Issue 11
Structure-Activity Relationship Studies on the Macrolide Exotoxin Mycolactone of Mycobacterium ulcerans
journal, March 2013
- Scherr, Nicole; Gersbach, Philipp; Dangy, Jean-Pierre
- PLoS Neglected Tropical Diseases, Vol. 7, Issue 3
Mycobacterium ulcerans toxic macrolide, mycolactone modulates the host immune response and cellular location of M. ulcerans in vitro and in vivo: Mycolactone and virulence of Mycobacterium ulcerans
journal, June 2005
- Adusumilli, Sarojini; Mve-Obiang, Armand; Sparer, Tim
- Cellular Microbiology, Vol. 7, Issue 9
Detection of Lipomannan in Cattle Infected with Bovine Tuberculosis
journal, January 2017
- Vu, Dung M.; Sakamuri, Rama M.; Waters, W. Ray
- Analytical Sciences, Vol. 33, Issue 4, p. 457-460
Determination of the Critical Concentration for Micelle Formation in Solutions of Colloidal Electrolytes by the Spectral Change of a Dye 1
journal, March 1947
- Corrin, M. L.; Harkins, William D.
- Journal of the American Chemical Society, Vol. 69, Issue 3
A Mycobacterium ulcerans Toxin, Mycolactone, Causes Apoptosis in Guinea Pig Ulcers and Tissue Culture Cells
journal, February 2000
- George, K. M.; Pascopella, L.; Welty, D. M.
- Infection and Immunity, Vol. 68, Issue 2
Mycolactones: immunosuppressive and cytotoxic polyketides produced by aquatic mycobacteria
journal, January 2008
- Hong, Hui; Demangel, Caroline; Pidot, Sacha J.
- Natural Product Reports, Vol. 25, Issue 3
Mycolactone Diffuses into the Peripheral Blood of Buruli Ulcer Patients - Implications for Diagnosis and Disease Monitoring
journal, July 2011
- Sarfo, Fred S.; Le Chevalier, Fabien; Aka, N'Guetta
- PLoS Neglected Tropical Diseases, Vol. 5, Issue 7
Photodegradation of the Mycobacterium ulcerans Toxin, Mycolactones: Considerations for Handling and Storage
journal, April 2012
- Marion, Estelle; Prado, Soizic; Cano, Camille
- PLoS ONE, Vol. 7, Issue 4
Characterization of Novel Macrolide Toxins, Mycolactones A and B, from a Human Pathogen, Mycobacterium ulcerans
journal, June 1999
- Gunawardana, Geewananda; Chatterjee, Delphi; George, Kathleen M.
- Journal of the American Chemical Society, Vol. 121, Issue 25
BCG-Mediated Protection against Mycobacterium ulcerans Infection in the Mouse
journal, March 2011
- Converse, Paul J.; Almeida, Deepak V.; Nuermberger, Eric L.
- PLoS Neglected Tropical Diseases, Vol. 5, Issue 3
FVB/N Mice Spontaneously Heal Ulcerative Lesions Induced by Mycobacterium ulcerans and Switch M. ulcerans into a Low Mycolactone Producer
journal, February 2016
- Marion, Estelle; Jarry, Ulrich; Cano, Camille
- The Journal of Immunology, Vol. 196, Issue 6
The Macrolide Toxin Mycolactone Promotes Bim-Dependent Apoptosis in Buruli Ulcer through Inhibition of mTOR
journal, March 2017
- Bieri, Raphael; Scherr, Nicole; Ruf, Marie-Thérèse
- ACS Chemical Biology, Vol. 12, Issue 5
Mycolactone Is Responsible for the Painlessness of Mycobacterium ulcerans Infection (Buruli Ulcer) in a Murine Study
journal, March 2008
- En, J.; Goto, M.; Nakanaga, K.
- Infection and Immunity, Vol. 76, Issue 5
Recent advances: role of mycolactone in the pathogenesis and monitoring of Mycobacterium ulcerans infection/Buruli ulcer disease : Mycolactone and pathogenesis of Buruli ulcer
journal, December 2015
- Sarfo, Fred Stephen; Phillips, Richard; Wansbrough-Jones, Mark
- Cellular Microbiology, Vol. 18, Issue 1
MYCOBACTERIUM ULCERANS INFECTION..: Clinical and Bacteriological Study of the First Cases Recognized in South East Asia.
journal, April 1966
- Pettit, John H. S.; Marchette, Nyven J.; Rees, R. J. W.
- British Journal of Dermatology, Vol. 78, Issue 4
Mycolactone: A Polyketide Toxin from Mycobacterium ulcerans Required for Virulence
journal, February 1999
- George, K. M.
- Science, Vol. 283, Issue 5403
Kinetics of mycolactone in human subcutaneous tissue during antibiotic therapy for Mycobacterium ulcerans disease
journal, April 2014
- Sarfo, Fred S.; Phillips, Richard O.; Zhang, Jihui
- BMC Infectious Diseases, Vol. 14, Issue 1
Total synthesis of mycolactones A and B
journal, June 2007
- Song, Fengbin; Fidanze, Steve; Benowitz, Andrew B.
- Tetrahedron, Vol. 63, Issue 26
Photochemistry of Mycolactone A/B, the Causative Toxin of Buruli Ulcer
journal, November 2012
- Xing, Yalan; Hande, Sudhir M.; Kishi, Yoshito
- Journal of the American Chemical Society, Vol. 134, Issue 46
Detection of Mycolactone A/B in Mycobacterium ulcerans–Infected Human Tissue
journal, January 2010
- Sarfo, Fred Stephen; Phillips, Richard O.; Rangers, Brian
- PLoS Neglected Tropical Diseases, Vol. 4, Issue 1
Total Syntheses of Mycolactone A/B and its Analogues for the Exploration of the Biology of Buruli Ulcer
journal, December 2017
- Saint-Auret, Sarah; Chany, Anne-Caroline; Casarotto, Virginie
- CHIMIA International Journal for Chemistry, Vol. 71, Issue 12
Rapid, Serial, Non-invasive Assessment of Drug Efficacy in Mice with Autoluminescent Mycobacterium ulcerans Infection
journal, December 2013
- Zhang, Tianyu; Li, Si-Yang; Converse, Paul J.
- PLoS Neglected Tropical Diseases, Vol. 7, Issue 12
Heterogeneity of Mycolactones Produced by Clinical Isolates of Mycobacterium ulcerans: Implications for Virulence
journal, February 2003
- Mve-Obiang, A.; Lee, R. E.; Portaels, F.
- Infection and Immunity, Vol. 71, Issue 2
Presentation matters: Impact of association of amphiphilic LPS with serum carrier proteins on innate immune signaling
journal, June 2018
- Stromberg, Loreen R.; Mendez, Heather M.; Kubicek-Sutherland, Jessica Z.
- PLOS ONE, Vol. 13, Issue 6
Association of lipoarabinomannan with high density lipoprotein in blood: Implications for diagnostics
journal, May 2013
- Sakamuri, Rama Murthy; Price, Dominique N.; Lee, Myungsun
- Tuberculosis, Vol. 93, Issue 3, p. 301-307
A Diverted Total Synthesis of Mycolactone Analogues: An Insight into Buruli Ulcer Toxins
journal, November 2011
- Chany, Anne-Caroline; Casarotto, Virginie; Schmitt, Marjorie
- Chemistry - A European Journal, Vol. 17, Issue 51
Mycolactone activation of Wiskott-Aldrich syndrome proteins underpins Buruli ulcer formation
journal, March 2013
- Guenin-Macé, Laure; Veyron-Churlet, Romain; Thoulouze, Maria-Isabel
- Journal of Clinical Investigation, Vol. 123, Issue 4
Membrane perturbing properties of toxin mycolactone from Mycobacterium ulcerans
journal, February 2018
- López, Cesar A.; Unkefer, Clifford J.; Swanson, Basil I.
- PLOS Computational Biology, Vol. 14, Issue 2
Distribution and Kinetics of Lipoprotein-Bound Lipoteichoic Acid
journal, June 2003
- Levels, J. H. M.; Abraham, P. R.; van Barreveld, E. P.
- Infection and Immunity, Vol. 71, Issue 6
Synthetic Variants of Mycolactone Bind and Activate Wiskott–Aldrich Syndrome Proteins
journal, August 2014
- Chany, Anne-Caroline; Veyron-Churlet, Romain; Tresse, Cédric
- Journal of Medicinal Chemistry, Vol. 57, Issue 17
The role of HDL in innate immunity
journal, October 2010
- Feingold, Kenneth R.; Grunfeld, Carl
- Journal of Lipid Research, Vol. 52, Issue 1
Chemistry of mycolactones, the causative toxins of Buruli ulcer
journal, March 2011
- Kishi, Y.
- Proceedings of the National Academy of Sciences, Vol. 108, Issue 17
Detection of Lipid and Amphiphilic Biomarkers for Disease Diagnostics
journal, July 2017
- Kubicek-Sutherland, Jessica; Vu, Dung; Mendez, Heather
- Biosensors, Vol. 7, Issue 4
Identification using LC-MSn of co-metabolites in the biosynthesis of the polyketide toxin mycolactone by a clinical isolate of Mycobacterium ulceransElectronic supplementary information (ESI) available: Experimental procedures and ESI-CID-MS/MS spectra of mycolactone and the five co-metabolites; MS3 spectrum of m/z 661 from the MS/MS of m/z 749; scheme showing the losses of mass 88 (C4H8O2) during the MS/MS of m/z 749 and the MS3 of m/z 661. See http://www.rsc.org/suppdata/cc/b3/b308163j/
journal, January 2003
- Hong, Hui; Gates, Paul J.; Staunton, James
- Chemical Communications, Issue 22
Uptake and cellular actions of mycolactone, a virulence determinant for Mycobacterium ulcerans
journal, February 2003
- Snyder, D. Scott; Small, P. L. C.
- Microbial Pathogenesis, Vol. 34, Issue 2
Distribution of Mycobacterium ulcerans in Buruli Ulcer Endemic and Non-Endemic Aquatic Sites in Ghana
journal, March 2008
- Williamson, Heather R.; Benbow, Mark E.; Nguyen, Khoa D.
- PLoS Neglected Tropical Diseases, Vol. 2, Issue 3
Mycolactone Diffuses from Mycobacterium ulcerans–Infected Tissues and Targets Mononuclear Cells in Peripheral Blood and Lymphoid Organs
journal, October 2008
- Hong, Hui; Coutanceau, Emmanuelle; Leclerc, Marion
- PLoS Neglected Tropical Diseases, Vol. 2, Issue 10
In vivo protection against endotoxin by plasma high density lipoprotein.
journal, December 1993
- Levine, D. M.; Parker, T. S.; Donnelly, T. M.
- Proceedings of the National Academy of Sciences, Vol. 90, Issue 24
HDL in sepsis – risk factor and therapeutic approach
journal, October 2015
- Morin, Emily E.; Guo, Ling; Schwendeman, Anna
- Frontiers in Pharmacology, Vol. 6
Highly sensitive, operationally simple, cost/time effective detection of the mycolactones from the human pathogen Mycobacterium ulcerans
journal, January 2010
- Spangenberg, Thomas; Kishi, Yoshito
- Chemical Communications, Vol. 46, Issue 9
Partial Purification and Characterization of Biological Effects of a Lipid Toxin Produced by Mycobacterium ulcerans
journal, February 1998
- George, Kathleen M.; Barker, Lucia P.; Welty, Diane M.
- Infection and Immunity, Vol. 66, Issue 2
Cytotoxic Activity of Mycobacterium ulcerans
journal, June 1974
- Read, John K.; Heggie, Charlotte M.; Meyers, Wayne M.
- Infection and Immunity, Vol. 9, Issue 6
A centrifugal microfluidic cross-flow filtration platform to separate serum from whole blood for the detection of amphiphilic biomarkers
journal, March 2021
- Lenz, Kiersten D.; Jakhar, Shailja; Chen, Jing W.
- Scientific Reports, Vol. 11, Issue 1
Structure-activity relationship studies on the macrolide exotoxin mycolactone of Mycobacterium ulcerans
text, January 2013
- Scherr, Nicole; Gersbach, Philipp; Dangy, Jean-Pierre
- Public Library of Science
Total Synthesis of Mycolactones A and B.
journal, October 2007
- Song, Fengbin; Fidanze, Steve; Benowitz, Andrew B.
- ChemInform, Vol. 38, Issue 42
The chemistry and biology of mycolactones
text, January 2017
- Gehringer, Matthias; Altmann, Karl-Heinz
- ETH Zurich
Structure-Activity Relationship Studies on the Macrolide Exotoxin Mycolactone of Mycobacterium ulcerans
text, January 2013
- Scherr, Nicole; Gersbach, Philipp; Dangy, Jean-Pierre
- ETH Zurich
Works referencing / citing this record:
Could Mycolactone Inspire New Potent Analgesics? Perspectives and Pitfalls
journal, September 2019
- Reynaert, Marie-Line; Dupoiron, Denis; Yeramian, Edouard
- Toxins, Vol. 11, Issue 9
Figures / Tables found in this record: