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Title: Impact of Fatty-Acid Labeling of Bacillus subtilis Membranes on the Cellular Lipidome and Proteome

Abstract

Developing cultivation methods that yield chemically and isotopically defined fatty acid (FA) compositions within bacterial cytoplasmic membranes establishes an in vivo experimental platform to study membrane biophysics and cell membrane regulation using novel approaches. Yet before fully realizing the potential of this method, it is prudent to understand the systemic changes in cells induced by the labeling procedure itself. In this work, analysis of cellular membrane compositions was paired with proteomics to assess how the proteome changes in response to the directed incorporation of exogenous FAs into the membrane of Bacillus subtilis. Key findings from this analysis include an alteration in lipid headgroup distribution, with an increase in phosphatidylglycerol lipids and decrease in phosphatidylethanolamine lipids, possibly providing a fluidizing effect on the cell membrane in response to the induced change in membrane composition. Changes in the abundance of enzymes involved in FA biosynthesis and degradation are observed; along with changes in abundance of cell wall enzymes and isoprenoid lipid production. The observed changes may influence membrane organization, and indeed the well-known lipid raft-associated protein flotillin was found to be substantially down-regulated in the labeled cells – as was the actin-like protein MreB. Taken as a whole, this study provides amore » greater depth of understanding for this important cell membrane experimental platform and presents a number of new connections to be explored in regard to modulating cell membrane FA composition and its effects on lipid headgroup and raft/cytoskeletal associated proteins.« less

Authors:
; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Laboratory Directed Research and Development (LDRD) Program; USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division
OSTI Identifier:
1618937
Alternate Identifier(s):
OSTI ID: 1649444
Grant/Contract Number:  
AC05-00OR22725; 6988; FWP ERKP752
Resource Type:
Published Article
Journal Name:
Frontiers in Microbiology
Additional Journal Information:
Journal Name: Frontiers in Microbiology Journal Volume: 11; Journal ID: ISSN 1664-302X
Publisher:
Frontiers Research Foundation
Country of Publication:
Switzerland
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Bacillus subtilis; biomembranes; proteomics; lipidomics; fatty-acids

Citation Formats

Nickels, Jonathan D., Poudel, Suresh, Chatterjee, Sneha, Farmer, Abigail, Cordner, Destini, Campagna, Shawn R., Giannone, Richard J., Hettich, Robert L., Myles, Dean A. A., Standaert, Robert F., Katsaras, John, and Elkins, James G. Impact of Fatty-Acid Labeling of Bacillus subtilis Membranes on the Cellular Lipidome and Proteome. Switzerland: N. p., 2020. Web. doi:10.3389/fmicb.2020.00914.
Nickels, Jonathan D., Poudel, Suresh, Chatterjee, Sneha, Farmer, Abigail, Cordner, Destini, Campagna, Shawn R., Giannone, Richard J., Hettich, Robert L., Myles, Dean A. A., Standaert, Robert F., Katsaras, John, & Elkins, James G. Impact of Fatty-Acid Labeling of Bacillus subtilis Membranes on the Cellular Lipidome and Proteome. Switzerland. doi:https://doi.org/10.3389/fmicb.2020.00914
Nickels, Jonathan D., Poudel, Suresh, Chatterjee, Sneha, Farmer, Abigail, Cordner, Destini, Campagna, Shawn R., Giannone, Richard J., Hettich, Robert L., Myles, Dean A. A., Standaert, Robert F., Katsaras, John, and Elkins, James G. Fri . "Impact of Fatty-Acid Labeling of Bacillus subtilis Membranes on the Cellular Lipidome and Proteome". Switzerland. doi:https://doi.org/10.3389/fmicb.2020.00914.
@article{osti_1618937,
title = {Impact of Fatty-Acid Labeling of Bacillus subtilis Membranes on the Cellular Lipidome and Proteome},
author = {Nickels, Jonathan D. and Poudel, Suresh and Chatterjee, Sneha and Farmer, Abigail and Cordner, Destini and Campagna, Shawn R. and Giannone, Richard J. and Hettich, Robert L. and Myles, Dean A. A. and Standaert, Robert F. and Katsaras, John and Elkins, James G.},
abstractNote = {Developing cultivation methods that yield chemically and isotopically defined fatty acid (FA) compositions within bacterial cytoplasmic membranes establishes an in vivo experimental platform to study membrane biophysics and cell membrane regulation using novel approaches. Yet before fully realizing the potential of this method, it is prudent to understand the systemic changes in cells induced by the labeling procedure itself. In this work, analysis of cellular membrane compositions was paired with proteomics to assess how the proteome changes in response to the directed incorporation of exogenous FAs into the membrane of Bacillus subtilis. Key findings from this analysis include an alteration in lipid headgroup distribution, with an increase in phosphatidylglycerol lipids and decrease in phosphatidylethanolamine lipids, possibly providing a fluidizing effect on the cell membrane in response to the induced change in membrane composition. Changes in the abundance of enzymes involved in FA biosynthesis and degradation are observed; along with changes in abundance of cell wall enzymes and isoprenoid lipid production. The observed changes may influence membrane organization, and indeed the well-known lipid raft-associated protein flotillin was found to be substantially down-regulated in the labeled cells – as was the actin-like protein MreB. Taken as a whole, this study provides a greater depth of understanding for this important cell membrane experimental platform and presents a number of new connections to be explored in regard to modulating cell membrane FA composition and its effects on lipid headgroup and raft/cytoskeletal associated proteins.},
doi = {10.3389/fmicb.2020.00914},
journal = {Frontiers in Microbiology},
number = ,
volume = 11,
place = {Switzerland},
year = {2020},
month = {5}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: https://doi.org/10.3389/fmicb.2020.00914

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