Incorporation of carbon and nitrogen atoms into proteins measured by protein-based stable isotope probing (Protein-SIP)

Rapid Communications in Mass Spectrometry : RCM
Nico JehmlichCarsten Vogt

Abstract

The identification of metabolically active microbial key players is fundamental for understanding the structure and functions of contaminant-degrading communities. The metabolic activity can be analysed by feeding the microbial culture with stable-isotope-labelled substrates and subsequently tracing their incorporation into the biomass. In this paper we present a method which is able to detect the incorporation of stable isotopes from the substrate into the proteins of a benzene-metabolising microorganism. Pseudomonas putida strain ML2 was grown under aerobic conditions with the substrates (12)C-benzene, (13)C-benzene or (15)N-ammonium and (12)C-benzene. Proteins of these cultures were resolved by two-dimensional gel electrophoresis (2-DE) and corresponding protein spots were subjected to matrix-assisted laser ionization/desorption mass spectrometric (MALDI-MS) analysis. The proteins of the (12)C-sample were identified by peptide mass fingerprinting (PMF) as well as by tandem mass spectrometric (MS/MS) measurements. The (13)C- or (15)N-content of the peptides from the labelling experiments was determined by MALDI-MS/MS. The incorporation of heavy isotopes into the proteins from cultures grown on (13)C-benzene and (15)N-ammonium...Continue Reading

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Citations

Apr 6, 2012·Applied Microbiology and Biotechnology·Sabine KleinsteuberCarsten Vogt
Dec 4, 2010·Nature Protocols·Nico JehmlichCarsten Vogt
Feb 3, 2011·Molecular & Cellular Proteomics : MCP·Chongle PanJillian F Banfield
Jan 28, 2014·Zhurnal evoliutsionnoĭ biokhimii i fiziologii·O V Matantseva, S O Skarlato
Aug 3, 2010·Applied Microbiology and Biotechnology·Nico JehmlichFrank Schmidt
Mar 19, 2013·Analytical and Bioanalytical Chemistry·Martin TaubertJana Seifert
Oct 2, 2012·Biotechnology Advances·Ondrej UhlikTomas Macek
May 15, 2015·Analytical Chemistry·Romain TrouillardIsabelle Ségalas-Milazzo
Feb 15, 2012·Geobiology·V Klepac-CerajA Pearson
Apr 27, 2011·Environmental Microbiology·Dominic BurgRicardo Cavicchioli
Apr 16, 2015·Microbial Biotechnology·Robert HeyerDirk Benndorf
Mar 13, 2015·FEMS Microbiology Ecology·Steffen KümmelCarsten Vogt
Apr 27, 2016·Current Opinion in Biotechnology·Nico JehmlichMartin von Bergen
Oct 12, 2011·International Archives of Allergy and Immunology·Sabine KespohlMonika Raulf-Heimsoth
May 25, 2010·FEMS Microbiology Ecology·Felipe BastidaHans H Richnow
Nov 13, 2010·Journal of Applied Microbiology·N JehmlichJ Seifert
Mar 17, 2012·Mass Spectrometry Reviews·Jana SeifertMartin von Bergen
Apr 23, 2014·Environmental Microbiology·Nicholas B JusticeJillian F Banfield
Jan 19, 2020·Applied and Environmental Microbiology·Laurey SteinkeDavid M Ward
Mar 28, 2019·Applied Microbiology and Biotechnology·Ayrat M ZiganshinSabine Kleinsteuber

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