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Title: MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine

Abstract

The cell envelope of Mycobacterium tuberculosis is notable for the abundance of mycolic acids (MAs), essential to mycobacterial viability, and of other species-specific lipids. The mycobacterial cell envelope is extremely hydrophobic, which contributes to virulence and antibiotic resistance. However, exactly how fatty acids and lipidic elements are transported across the cell envelope for cell-wall biosynthesis is unclear. Mycobacterial membrane protein Large 3 (MmpL3) is essential and required for transport of trehalose monomycolates (TMMs), precursors of MA-containing trehalose dimycolates (TDM) and mycolyl arabinogalactan peptidoglycan, but the exact function of MmpL3 remains elusive. Here, we report a crystal structure of Mycobacterium smegmatis MmpL3 at a resolution of 2.59 Å, revealing a monomeric molecule that is structurally distinct from all known bacterial membrane proteins. A previously unknown MmpL3 ligand, phosphatidylethanolamine (PE), was discovered inside this transporter. We also show, via native mass spectrometry, that MmpL3 specifically binds both TMM and PE, but not TDM, in the micromolar range. These observations provide insight into the function of MmpL3 and suggest a possible role for this protein in shuttling a variety of lipids to strengthen the mycobacterial cell wall.

Authors:
 [1];  [1]; ORCiD logo [2];  [3]; ORCiD logo [2]; ORCiD logo [1]
  1. Case Western Reserve Univ., Cleveland, OH (United States)
  2. Univ. of Oxford (United Kingdom)
  3. Oregon Health & Science Univ., Portland, OR (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1531017
Report Number(s):
[AC02-06CH11357]
[Journal ID: ISSN 0027-8424]
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
[ Journal Volume: 116; Journal Issue: 23]; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; mycobacterial membrane protein Large; cell-wall biogenesis; MmpL3 transporter; X-ray crystallography; native mass spectrometry

Citation Formats

Su, Chih-Chia, Klenotic, Philip A., Bolla, Jani Reddy, Purdy, Georgiana E., Robinson, Carol V., and Yu, Edward W. MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine. United States: N. p., 2019. Web. doi:10.1073/pnas.1901346116.
Su, Chih-Chia, Klenotic, Philip A., Bolla, Jani Reddy, Purdy, Georgiana E., Robinson, Carol V., & Yu, Edward W. MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine. United States. doi:10.1073/pnas.1901346116.
Su, Chih-Chia, Klenotic, Philip A., Bolla, Jani Reddy, Purdy, Georgiana E., Robinson, Carol V., and Yu, Edward W. Tue . "MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine". United States. doi:10.1073/pnas.1901346116.
@article{osti_1531017,
title = {MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine},
author = {Su, Chih-Chia and Klenotic, Philip A. and Bolla, Jani Reddy and Purdy, Georgiana E. and Robinson, Carol V. and Yu, Edward W.},
abstractNote = {The cell envelope of Mycobacterium tuberculosis is notable for the abundance of mycolic acids (MAs), essential to mycobacterial viability, and of other species-specific lipids. The mycobacterial cell envelope is extremely hydrophobic, which contributes to virulence and antibiotic resistance. However, exactly how fatty acids and lipidic elements are transported across the cell envelope for cell-wall biosynthesis is unclear. Mycobacterial membrane protein Large 3 (MmpL3) is essential and required for transport of trehalose monomycolates (TMMs), precursors of MA-containing trehalose dimycolates (TDM) and mycolyl arabinogalactan peptidoglycan, but the exact function of MmpL3 remains elusive. Here, we report a crystal structure of Mycobacterium smegmatis MmpL3 at a resolution of 2.59 Å, revealing a monomeric molecule that is structurally distinct from all known bacterial membrane proteins. A previously unknown MmpL3 ligand, phosphatidylethanolamine (PE), was discovered inside this transporter. We also show, via native mass spectrometry, that MmpL3 specifically binds both TMM and PE, but not TDM, in the micromolar range. These observations provide insight into the function of MmpL3 and suggest a possible role for this protein in shuttling a variety of lipids to strengthen the mycobacterial cell wall.},
doi = {10.1073/pnas.1901346116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = [23],
volume = [116],
place = {United States},
year = {2019},
month = {5}
}

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