scholarly journals MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine

2019 ◽  
Author(s):  
Chih-Chia Su ◽  
Philip Klenotic ◽  
Jani Reddy Bolla ◽  
Georgiana Purdy ◽  
Carol Robinson ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Envelope ◽  
Native Mass Spectrometry ◽  
Mycolic Acids ◽  
Mycobacterial Cell ◽  
Exact Function ◽  
Species Specific ◽  
Mycobacterial Cell Wall ◽  
Trehalose Monomycolate ◽  
Lipid Transporter

The cell envelope of Mycobacterium tuberculosis is notable for the abundance of mycolic acids (MAs), which are essential to mycobacterial viability, and other species-specific lipids. The mycobacterial cell envelope is extremely hydrophobic, contributes to virulence and antibiotic resistance. Yet, 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 (mAGP), but the exact function of MmpL3 remains elusive. Here, we report a high-resolution crystal structure of M. smegmatis MmpL3, 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.

scholarly journals MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine

2019 ◽  
Vol 116 (23) ◽  
pp. 11241-11246 ◽  
Author(s):  
Chih-Chia Su ◽  
Philip A. Klenotic ◽  
Jani Reddy Bolla ◽  
Georgiana E. Purdy ◽  
Carol V. Robinson ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Envelope ◽  
Native Mass Spectrometry ◽  
Mycolic Acids ◽  
Mycobacterial Cell ◽  
Exact Function ◽  
Species Specific ◽  
Mycobacterial Cell Wall ◽  
Trehalose Monomycolate ◽  
Lipid Transporter

The cell envelope ofMycobacterium tuberculosisis 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 ofMycobacterium smegmatisMmpL3 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.

scholarly journals Synthesis of a Di-Mycoloyl Tri-Arabinofuranosyl Glycerol Fragment of the Mycobacterial Cell Wall, Based on Synthetic Mycolic Acids

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3596
Author(s):  
Ali ◽  
Mohammed ◽  
Dulayymi ◽  
Baird
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Stereoselective Synthesis ◽  
Complex Mixtures ◽  
Mycolic Acids ◽  
Mycobacterial Cell ◽  
Antigenic Properties ◽  
Mycobacterial Cell Wall ◽  
Structural Classes

Fragments of mycobacterial cell walls such as arabinoglycerol mycolate and dimycoloyl diarabinoglycerol, comprising complex mixtures of mycolic acids, have immunostimulatory and antigenic properties. A related di-mycoloyl tri-arabinofuranosyl glycerol fragment has been isolated from cell wall hydrolysates. An effective stereoselective synthesis of tri-arabinofuranosyl glycerol, followed by coupling with stereochemically defined mycolic acids of different structural classes, to provide unique di-mycoloyl tri-arabinofuranosyl glycerols is now described.

scholarly journals Mycobacteriophage Ms6 LysB specifically targets the outer membrane of Mycobacterium smegmatis

Microbiology ◽  
2010 ◽  
Vol 156 (5) ◽  
pp. 1497-1504 ◽  
Author(s):  
Filipa Gil ◽  
Anna E. Grzegorzewicz ◽  
Maria João Catalão ◽  
João Vital ◽  
Michael R. McNeil ◽  
...  
Keyword(s):  
Cell Wall ◽  
Outer Membrane ◽  
Mycobacterium Smegmatis ◽  
Mycolic Acid ◽  
Head Group ◽  
Lipolytic Enzyme ◽  
Ester Bond ◽  
Mycolic Acids ◽  
Mycobacterial Cell ◽  
Mycobacterial Cell Wall

LysB, a mycobacteriophage Ms6-encoded protein, was previously identified as a lipolytic enzyme able to hydrolyse the ester bond in lipase and esterase substrates. In the present work, we show that LysB can hydrolyse lipids containing mycolic acids from the outer membrane of the mycobacterial cell wall. LysB was shown to hydrolyse the mycolic acids from the mycolyl-arabinogalactan–peptidoglycan complex where the mycolates of the inner leaflet of the outer membrane are covalently attached to an arabinosyl head group. In addition, treatment of the extractable lipids from Mycobacterium smegmatis, Mycobacterium bovis BCG and Mycobacterium tuberculosis H37Ra with LysB showed that trehalose 6,6′-dimycolate (TDM), a trehalose diester of two mycolic acid molecules, was hydrolysed by the enzyme. We have also determined the structures of the mycolic acid molecules that form the M. smegmatis TDM. The identification of a phage-encoded enzyme that targets the outer membrane of the mycobacterial cell wall enhances our understanding of the mechanism of mycobacteriophage lysis.

ChemInform Abstract: The Mycobacterial Cell-Wall as Target for Antimycobacterial Drugs. Part 1. Synthesis and Activity of Some Diphenylalkyl Analogues of Mycolic Acids.

ChemInform ◽  
1988 ◽  
Vol 19 (20) ◽  
Author(s):  
D. C. LEYSEN ◽  
A. HAEMERS ◽  
L. BLANCHAERT ◽  
I. VAN ASSCHE ◽  
W. BOLLAERT ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mycolic Acids ◽  
Mycobacterial Cell ◽  
Mycobacterial Cell Wall

scholarly journals Lcp1 Is a Phosphotransferase Responsible for Ligating Arabinogalactan to Peptidoglycan in Mycobacterium tuberculosis

mBio ◽  
2016 ◽  
Vol 7 (4) ◽  
Author(s):  
James Harrison ◽  
Georgina Lloyd ◽  
Maju Joe ◽  
Todd L. Lowary ◽  
Edward Reynolds ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mycobacterium Tuberculosis ◽  
Essential Gene ◽  
Cell Envelope ◽  
New Drugs ◽  
Cell Wall Assembly ◽  
Mycobacterial Cell ◽  
Unique Cell ◽  
Mycobacterial Cell Wall ◽  
Wall Assembly

ABSTRACT Mycobacterium tuberculosis , the etiological agent of tuberculosis (TB), has a unique cell envelope which accounts for its unusual low permeability and contributes to resistance against common antibiotics. The main structural elements of the cell wall consist of a cross-linked network of peptidoglycan (PG) in which some of the muramic acid residues are covalently attached to a complex polysaccharide, arabinogalactan (AG), via a unique α- l -rhamnopyranose–(1→3)-α- d -GlcNAc-(1→P) linker unit. While the molecular genetics associated with PG and AG biosynthetic pathways have been largely delineated, the mechanism by which these two major pathways converge has remained elusive. In Gram-positive organisms, the LytR-CpsA-Psr (LCP) family of proteins are responsible for ligating cell wall teichoic acids to peptidoglycan, through a linker unit that bears a striking resemblance to that found in mycobacterial arabinogalactan. In this study, we have identified Rv3267 as a mycobacterial LCP homolog gene that encodes a phosphotransferase which we have named Lcp1. We demonstrate that lcp1 is an essential gene required for cell viability and show that recombinant Lcp1 is capable of ligating AG to PG in a cell-free radiolabeling assay. IMPORTANCE Tuberculosis is an infectious disease caused by the bacterial organism Mycobacterium tuberculosis . Survival of M. tuberculosis rests critically on the integrity of its unique cell wall; therefore, a better understanding of how the genes and enzymes involved in cell wall assembly work is fundamental for us to develop new drugs to treat this disease. In this study, we have identified Lcp1 as an essential phosphotransferase that ligates together arabinogalactan and peptidoglycan, two crucial cell wall macromolecules found within the mycobacterial cell wall. The discovery of Lcp1 sheds new light on the final stages of mycobacterial cell wall assembly and represents a key biosynthetic step that could be exploited for new anti-TB drug discovery.

scholarly journals Structures of the mycobacterial membrane protein MmpL3 reveal its mechanism of lipid transport

PLoS Biology ◽  
2021 ◽  
Vol 19 (8) ◽  
pp. e3001370
Author(s):  
Chih-Chia Su ◽  
Philip A. Klenotic ◽  
Meng Cui ◽  
Meinan Lyu ◽  
Christopher E. Morgan ◽  
...  
Keyword(s):  
Cell Wall ◽  
Membrane Protein ◽  
Md Simulations ◽  
Mycolic Acid ◽  
Lipid Transport ◽  
X Ray Crystallography ◽  
Trehalose Dimycolate ◽  
Mycobacterial Cell ◽  
Mycobacterial Cell Wall ◽  
Trehalose Monomycolate

The mycobacterial membrane protein large 3 (MmpL3) transporter is essential and required for shuttling the lipid trehalose monomycolate (TMM), a precursor of mycolic acid (MA)-containing trehalose dimycolate (TDM) and mycolyl arabinogalactan peptidoglycan (mAGP), in Mycobacterium species, including Mycobacterium tuberculosis and Mycobacterium smegmatis. However, the mechanism that MmpL3 uses to facilitate the transport of fatty acids and lipidic elements to the mycobacterial cell wall remains elusive. Here, we report 7 structures of the M. smegmatis MmpL3 transporter in its unbound state and in complex with trehalose 6-decanoate (T6D) or TMM using single-particle cryo-electron microscopy (cryo-EM) and X-ray crystallography. Combined with calculated results from molecular dynamics (MD) and target MD simulations, we reveal a lipid transport mechanism that involves a coupled movement of the periplasmic domain and transmembrane helices of the MmpL3 transporter that facilitates the shuttling of lipids to the mycobacterial cell wall.

Conserved Molecular Superlattices in a Series of Homologous Synthetic Mycobacterial Cell-Wall Lipids Forming Interdigitated Bilayers

Langmuir ◽  
2016 ◽  
Vol 32 (48) ◽  
pp. 12693-12701 ◽  
Author(s):  
Birte Martin-Bertelsen ◽  
Anan Yaghmur ◽  
Henrik Franzyk ◽  
Sarah Justesen ◽  
Jacob J. K. Kirkensgaard ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mycobacterial Cell ◽  
Mycobacterial Cell Wall
Sign in / Sign up

Export Citation Format

Share Document