scholarly journals Dewetting-induced formation and mechanical properties of synthetic bacterial outer membrane models (GUVs) with controlled inner-leaflet lipid composition

Soft Matter ◽  
2019 ◽  
Vol 15 (19) ◽  
pp. 3938-3948 ◽  
Author(s):  
Sepehr Maktabi ◽  
Jeffrey W. Schertzer ◽  
Paul R. Chiarot

We report on a microfluidic technique for fabricating monodisperse asymmetric giant unilamellar vesicles (GUVs) possessing the Gram-negative bacterial outer membrane lipid composition.

2011 ◽  
Vol 286 (15) ◽  
pp. 12850-12859 ◽  
Author(s):  
Matthias Keck ◽  
Nicolas Gisch ◽  
Hermann Moll ◽  
Frank-Jörg Vorhölter ◽  
Klaus Gerth ◽  
...  

Langmuir ◽  
2014 ◽  
Vol 31 (1) ◽  
pp. 404-412 ◽  
Author(s):  
Luke A. Clifton ◽  
Maximilian W. A. Skoda ◽  
Anton P. Le Brun ◽  
Filip Ciesielski ◽  
Ivan Kuzmenko ◽  
...  

2009 ◽  
Vol 191 (23) ◽  
pp. 7323-7332 ◽  
Author(s):  
Xavier Meniche ◽  
Cécile Labarre ◽  
Célia de Sousa-d'Auria ◽  
Emilie Huc ◽  
Françoise Laval ◽  
...  

ABSTRACT Corynebacterineae are gram-positive bacteria that possess a true outer membrane composed of mycolic acids and other lipids. Little is known concerning the modulation of mycolic acid composition and content in response to changes in the bacterial environment, especially temperature variations. To address this question, we investigated the function of the Rv3802c gene, a gene conserved in Corynebacterineae and located within a gene cluster involved in mycolic acid biosynthesis. We showed that the Rv3802 ortholog is essential in Mycobacterium smegmatis, while its Corynebacterium glutamicum ortholog, NCgl2775, is not. We provided evidence that the NCgl2775 gene is transcriptionally induced under heat stress conditions, and while the corresponding protein has no detectable activity under normal growth conditions, the increase in its expression triggers an increase in mycolic acid biosynthesis concomitant with a decrease in phospholipid content. We demonstrated that these lipid modifications are part of a larger outer membrane remodeling that occurs in response to exposure to a moderately elevated temperature (42°C). In addition to showing an increase in the ratio of saturated corynomycolates to unsaturated corynomycolates, our results strongly suggested that the balance between mycolic acids and phospholipids is modified inside the outer membrane following a heat challenge. Furthermore, we showed that these lipid modifications help the bacteria to protect against heat damage. The NCgl2775 protein and its orthologs thus appear to be a protein family that plays a role in the regulation of the outer membrane lipid composition of Corynebacterineae under stress conditions. We therefore propose to name this protein family the envelope lipids regulation factor (ElrF) family.


2013 ◽  
Vol 10 (89) ◽  
pp. 20130810 ◽  
Author(s):  
Luke A. Clifton ◽  
Maximilian W. A. Skoda ◽  
Emma L. Daulton ◽  
Arwel V. Hughes ◽  
Anton P. Le Brun ◽  
...  

The Gram-negative bacterial outer membrane (OM) is a complex and highly asymmetric biological barrier but the small size of bacteria has hindered advances in in vivo examination of membrane dynamics. Thus, model OMs, amenable to physical study, are important sources of data. Here, we present data from asymmetric bilayers which emulate the OM and are formed by a simple two-step approach. The bilayers were deposited on an SiO 2 surface by Langmuir–Blodgett deposition of phosphatidylcholine as the inner leaflet and, via Langmuir–Schaefer deposition, an outer leaflet of either Lipid A or Escherichia coli rough lipopolysaccharides (LPS). The membranes were examined using neutron reflectometry (NR) to examine the coverage and mixing of lipids between the bilayer leaflets. NR data showed that in all cases, the initial deposition asymmetry was mostly maintained for more than 16 h. This stability enabled the sizes of the headgroups and bilayer roughness of 1,2-dipalmitoyl- sn -glycero-3-phosphocholine and Lipid A, Rc-LPS and Ra-LPS to be clearly resolved. The results show that rough LPS can be manipulated like phospholipids and used to fabricate advanced asymmetric bacterial membrane models using well-known bilayer deposition techniques. Such models will enable OM dynamics and interactions to be studied under in vivo -like conditions.


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