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Probing the Subcellular Localization of Hopanoid Lipids in Bacteria Using NanoSIMS

Doughty, David M. and Dieterle, Michael and Sessions, Alex L. and Fischer, Woodward W. and Newman, Dianne K. (2014) Probing the Subcellular Localization of Hopanoid Lipids in Bacteria Using NanoSIMS. PLoS ONE, 9 (1). Art. No. e84455. ISSN 1932-6203. PMCID PMC3883690. http://resolver.caltech.edu/CaltechAUTHORS:20140206-114311185

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Abstract

The organization of lipids within biological membranes is poorly understood. Some studies have suggested lipids group into microdomains within cells, but the evidence remains controversial due to non-native imaging techniques. A recently developed NanoSIMS technique indicated that sphingolipids group into microdomains within membranes of human fibroblast cells. We extended this NanoSIMS approach to study the localization of hopanoid lipids in bacterial cells by developing a stable isotope labeling method to directly detect subcellular localization of specific lipids in bacteria with ca. 60 nm resolution. Because of the relatively small size of bacterial cells and the relative abundance of hopanoid lipids in membranes, we employed a primary ^2H-label to maximize our limit of detection. This approach permitted the analysis of multiple stable isotope labels within the same sample, enabling visualization of subcellular lipid microdomains within different cell types using a secondary label to mark the growing end of the cell. Using this technique, we demonstrate subcellular localization of hopanoid lipids within alpha-proteobacterial and cyanobacterial cells. Further, we provide evidence of hopanoid lipid domains in between cells of the filamentous cyanobacterium Nostoc punctiforme. More broadly, our method provides a means to image lipid microdomains in a wide range of cell types and test hypotheses for their functions in membranes.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1371/journal.pone.0084455DOIArticle
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0084455PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3883690/PubMed CentralArticle
ORCID:
AuthorORCID
Sessions, Alex L.0000-0001-6120-2763
Fischer, Woodward W.0000-0002-8836-3054
Newman, Dianne K.0000-0003-1647-1918
Additional Information:© 2014 Doughty et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received August 13, 2013; Accepted November 22, 2013; Published January 7, 2014. Grants to D.K.N. from the Howard Hughes Medical Institute (HHMI) and the NASA Exobiology program (NNX12AD93G) supported this work. D.K.N. is an HHMI Investigator. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Dr. Chia-Hung Wu for generating mutant strains of R. palustris and assistance with mass spectrometric analysis and Dr. Gargi Kulkarni for help with mCherry fusion strain construction. We thank Dr. Yungbin Guan, Prof. John Eiler and the Center for Microanalysis at the California Institute of Technology for assistance with NanoSIMS.
Funders:
Funding AgencyGrant Number
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NASANNX12AD93G
PubMed Central ID:PMC3883690
Record Number:CaltechAUTHORS:20140206-114311185
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20140206-114311185
Official Citation:Doughty DM, Dieterle M, Sessions AL, Fischer WW, Newman DK (2014) Probing the Subcellular Localization of Hopanoid Lipids in Bacteria Using NanoSIMS. PLoS ONE 9(1): e84455. doi:10.1371/journal.pone.0084455
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:43703
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:06 Feb 2014 22:49
Last Modified:24 Oct 2017 16:21

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