Here we describe a novel surface sampling technique termed pressurized liquid extraction surface analysis (PLESA), which in combination with a dedicated high resolution shotgun lipidomics routine enables both quantification and in-depth structural characterization of molecular lipid species extracted directly from tissue sections.
A: The laboratory is split 50/50 between proteomics and lipidomics research. While working on protein analysis, such as identifying protein interaction networks or characterizing the proteomes of organisms that are related very distantly to organisms with sequenced genomes, we also attempt to better quantify the lipidome of various organelles, cells and tissues.
Q: WHAT ARE YOUR PRIMARY GOALS?
A: In lipidomics, we forge the alliance with developmental biology. The primary goal of the group is to combine lipidomics with developmental biology. As organisms grow and develop from a single cell, newly differentiated tissues require their own unique membrane lipid composition. We hope to characterize these tailored changes to better understand how inherited defects in lipid metabolism cause disease. We are equally interested in lipidomes of membrane microdomains and the biological significance of its remarkable complexity.
Q: WHY DID YOU INCORPORATE THE TRIVERSA NANOMATE® INTO YOUR LABORATORY?
A: We had a need for automated nanoflow direct-infusion capabilities. Shotgun lipidomics relies on low and stable flow rates, and the TriVersa NanoMate® has this demonstrated ability. We have purchased three additional instruments because they allow us to rapidly switch between lipids and proteomic analysis.
Q: DO YOU HAVE ANY PUBLICATIONS OF PRESENTATIONS USING THE TRIVERSA NANOMATE®? Publication Highlight 2021: Hormone-Sensitive Lipase Couples Intergenerational Sterol Metabolism to Reproductive Success Christoph Heier Is a corresponding author , Oskar Knittelfelder, Harald F Hofbauer, Wolfgang Mende, Ingrid Pörnbacher, Laura Schiller, Gabriele Schoiswohl, Hao Xie, Sebastian Grönke, Andrej Shevchenko, Ronald P Kühnlein
Hormone-sensitive lipase (Hsl) was identified as an ancestral regulator of SE degradation, which improves intergenerational sterol transfer and reproductive success in flies.
Other Publications:
Knittelfelder, O., Prince, E., Sales, S., Fritzsche, E., Woehner, T., Brankatschk, M., Shevchenko, A. (2020) Sterols as dietary markers for Drosophila melanogaster. BBA – Molecular and Cell Biology of Lipids, 1865 (7), 1388-1981. DOI: 10.1016/j.bbalip.2020.158683
Trautenberg, L.C., Knittelfelder, O., Hofmann, C., Shevchenko, A., Brankatschk, M., Prince, E. (2020) How to use the development of individual Drosophila larvae as a metabolic sensor. Journal of Insect Biology, 126, 0022-1910. DOI: 10.1016/j.jinsphys.2020.104095
Wang, Y., Hinz, S., Uckermann, O., Hoenscheid, P., von Schoenfels, W., Burmesiter, G., Hendricks, A., Ackerman, J.M., Baretton, G.B., Hampe, J., Brosch, M., Schafmayer, C., Shevchenko, A., Seissig, S. (2020) Shotgun lipidomics-based characterization of the landscape of lipid metabolism in colorectal cancer. BBA – Molecular and Cell Biology of Lipids, 1865 (3), 1388-1981. DOI: 10.1016/j.bbalip.2019.158579
Finkelstein, S. Gospe III, S.M., Schuhmann, K., Shevkenko, A., Arshavsky, V.M., Lobanova, E.S. (2020) Phophoinositide profile of the mouse retina. Cells 9(6), 1417. DOI: 10.3390/cells9061417
Brankatschk, M., Gutmann, T., Knittelfelder, O., Palladini, A., Prince, E., Grzybek, M., Brankatschk, B., Shevchenko, A., Coskun, U., Eaton, S. (2018) A temperature-dependent switch in feeding preference improves drosphila development and survival in the cold. Developmental Cell, 46, 6, 781-793.e4. DOI: 10.1016/j.devcel.2018.05.028
Fernandez, C., Sandin, M., Sampaio, J.L., Almgren, P., Narkiewicz, K., Hoffmann, M., Hedner, T., Wahlstrand, B., Simons, K., Shevchenko, A., James, P., Melander, O. (2013) Plasma lipid composition and risk of developing cardiovascular disease. PLOS One. DOI: 10.1371/journal.pone.0071846
Ghosh, A., Kling, T., Snaidero, N., Sampaio, J.L., Shevchenko, A., Gras, H., Geurten, B., Goepfert, M.C., Schulz, J.B., Voigt, A., Simons, M. (2013) A global in vivo drosophila RNAi screen identifies a key role of ceramide phosphothanolamine for glial ensheathment of axons. PLOS Genetics. DOI: 10.1371/journal.pgen.1003980
Schuhmann, K., Almeida, R., Baumert, M., Herzog, R., Bornstein, S.R. and Shevchenko, A. (2012) Shotgun lipidomics on a LTQ Orbitrap mass spectrometer by successive switching between acquisition polarity modes. J. Mass. Spectrom., 47: 96-104. DOI: 10.1002/jms.2031
Carvalho, M., Sampaio, J.L., Palm, W., Brankatschk, M., Eaton, S., Shevchenko, A. (2012) Effects of diet and development on the Drosophila lipidome. Mol Syst Biol, 8:600. DOI: 10.1038/msb.2012.29
Luerschner, L. Richter, D., Hannibal-Bach, H.K., Gaebler, A., Shevchenko, A., Ejsing, C.S., Thiele, C. (2012) Exogenous ether lipids predominantly target mitochondria. PLOS ONE. DOI: 10.1371/journal.pone.0031342
Sampaio, J.L., Gerl, M.J., Klose, C., Ejsing, C.S., Beug, H., Simons, K., Shevchenko, A. (2011) Membrane lipidome of an epithelial cell line. PNAS, 108 (5), 1903-1907. DOI: 10.1073/pnas.1019267108
Ejsing, C.S., Sampaio, J.L., Surendranath, V., Duchoslav, E., Ekroos, K., Klemm, R.W., Simons, K., Shevchenko, A. (2009) Global analysis of the yeast lipidome by quantitative shotgun mass spectrometry. PNAS, 106 (7), 2136-2141. DOI: 10.1073/pnas.0811700106
In this webinar, Dr. Han describes how he uses the TriVersa NanoMate’s chip-based nanoelectrospray ionization capabilities in infusion mode to obtain more information from complex samples and faster lipids analysis with no sample-to-sample carryover. The interests of Dr. Han’s laboratory have been focused on the altered lipid metabolism, trafficking, and homeostasis under patho(physio)logical conditions. Currently, there are three specific areas explored in his laboratory including (1) extension of the shotgun lipidomics technology for increased penetrance into the low abundance regime of a cellular lipidome with emphasis on high throughput, and bioinformatics; (2) investigation of the biochemical mechanisms underlying the altered lipid content and composition in metabolic syndrome; and (3) identification of the biochemical mechanisms responsible for the sulfatide depletion and ceramide elevation at the very earliest stages of Alzheimer’s disease.
Presented by: Xianlin Han, Ph.D., Professor, Diabetes and Obesity Research Center, Sanford-Burnham Medical Research Institute
Presented by: Gavin Reid, Associate Professor at Michigan State University
A large number of studies have demonstrated that disruption of lipid metabolism or signaling pathways can play a key role in the onset and progression of human disease, including cancer and diabetes. Thus, a comparative analysis of changes in individual lipids or lipid profiles (i.e., the lipidome) between normal and diseased cells, tissues, organs, or accessible bodily fluids (e.g., tumor interstitial fluid, blood plasma or serum), may enable the identification and characterization of lipids that can serve as effective biomarker signatures of the disease. In this presentation, the development and application of a straightforward and high throughout analysis strategy consisting of high-resolution ‘shotgun’ mass spectrometry (MS), ‘targeted’ tandem mass spectrometry (MS/MS), functional group specific chemical modification and in situ liquid extraction of cell culture samples is described for the comprehensive identification, characterization and quantification of multiple lipid classes from within a colon adenocarcinoma cell line, SW480, and its metastasized derivative, SW620.
The rapid identification of radiolabeled compounds would clearly be beneficial for applied clinical and preclinical tracers used for imaging with positron emission tomography (PET). The limited uptake of mass spectrometry in the imaging field has been due to several factors including concerns regarding adequate sensitivity, instrument footprint in the highly constrained space requirements for the majority of labs and high initial capital costs plus high ongoing maintenance costs.
The introduction of the expression Compact Mass Spectrometer (CMS) at the 2012 Spring ACS National Meeting addressed for the first time three of the concerns: space, capital cost and maintenance costs. Only the question of adequate sensitivity remained to be answered.
