Next Article in Journal
A New Topical Eye Drop Containing LyeTxI-b, A Synthetic Peptide Designed from A Lycosa erithrognata Venom Toxin, Was Effective to Treat Resistant Bacterial Keratitis
Next Article in Special Issue
Recombinant Antibodies against Mycolactone
Previous Article in Journal
A New Method for Extraction and Analysis of Ricin Samples through MALDI-TOF-MS/MS
Previous Article in Special Issue
Molecular Docking and Dynamics Simulation Studies Predict Munc18b as a Target of Mycolactone: A Plausible Mechanism for Granule Exocytosis Impairment in Buruli Ulcer Pathogenesis
Article Menu
Issue 4 (April) cover image

Export Article

Open AccessArticle

Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone

1
Chemistry Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA
2
Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA
3
Department of Medicine, Johns Hopkins University Center for Tuberculosis Research, Baltimore, MD 21218, USA
*
Author to whom correspondence should be addressed.
Toxins 2019, 11(4), 202; https://doi.org/10.3390/toxins11040202
Received: 30 January 2019 / Revised: 26 February 2019 / Accepted: 1 April 2019 / Published: 4 April 2019
(This article belongs to the Special Issue Mycolactone: Lipid-Like Immunosuppressive Toxin of Buruli Ulcer)
  |  
PDF [1296 KB, uploaded 4 April 2019]
  |     |  

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 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. View Full-Text
Keywords: mycolactone; storage and handling; amphiphilic; lipoproteins mycolactone; storage and handling; amphiphilic; lipoproteins
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Kubicek-Sutherland, J.Z.; Vu, D.M.; Anderson, A.S.; Sanchez, T.C.; Converse, P.J.; Martí-Arbona, R.; Nuermberger, E.L.; Swanson, B.I.; Mukundan, H. Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone. Toxins 2019, 11, 202.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Toxins EISSN 2072-6651 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top