Exposure to natural and anthropogenic compounds can potentially alter the proteome in body fluids and tissues of living organisms, and by applying proteomics it is possible to discover, identify and understand such alterations. This study show results from a proteomic approach where one- or multidimensional separation (MudPIT) combined with high-accuracy tandem mass spectrometry (i.e. LTQ Orbitrap) were used to identify proteins from a non-model organism (Salmo salar). An optimized two-dimensional method resulted in more than 680 proteins identified with high significance compared to 197 proteins identified using a one-dimensional separation. Thus, MudPIT proteomics greatly increase the number of successful protein identification studies in ecotoxicology, and could potentially provide more insight into chemical modes of actions.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.marenvres.2010.03.008 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Differential Precipitation of Proteins: A Simple Protein Fractionation Strategy to Gain Biological Insights with Proteomics.
J Am Soc Mass Spectrom
September 2023
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California 92037, United States.
Differential precipitation of proteins (DiffPOP) is a simple technique for fractionating complex protein mixtures. Using stepwise addition of acidified methanol, ten distinct subsets of proteins can be selectively precipitated by centrifugation and identified by mass spectrometry-based proteomics. We have previously shown that the ability of a protein to resist precipitation can be altered by drug binding, which enabled us to identify a novel drug-target interaction.
View Article and Find Full Text PDFParallel Channels-Multidimensional Protein Identification Technology.
J Am Soc Mass Spectrom
July 2020
Center for Precision Medicine Multi-omics Research, Peking University Health Science Center, Beijing 100191, China.
Multidimensional protein identification (MudPIT), developed in the Yates Laboratory 20 years ago, is regarded as a powerful tool for proteomics research. Due to its remarkable online separation advantages, MudPIT has been widely used to facilitate discoveries in the field of proteomics research. However, it has one major disadvantage: the process of eluting peptides during strong cation exchange introduces salts, of different concentrations, into the mass spectrometer.
View Article and Find Full Text PDFA Robust and Universal Metaproteomics Workflow for Research Studies and Routine Diagnostics Within 24 h Using Phenol Extraction, FASP Digest, and the MetaProteomeAnalyzer.
Front Microbiol
August 2019
Bioprocess Engineering, Otto von Guericke University Magdeburg, Magdeburg, Germany.
The investigation of microbial proteins by mass spectrometry (metaproteomics) is a key technology for simultaneously assessing the taxonomic composition and the functionality of microbial communities in medical, environmental, and biotechnological applications. We present an improved metaproteomics workflow using an updated sample preparation and a new version of the MetaProteomeAnalyzer software for data analysis. High resolution by multidimensional separation (GeLC, MudPIT) was sacrificed to aim at fast analysis of a broad range of different samples in less than 24 h.
View Article and Find Full Text PDFIsolation, Proteomic Analysis, and Microscopy Confirmation of the Liver Nuclear Envelope Proteome.
Methods Mol Biol
December 2017
The Wellcome Trust Centre for Cell Biology and Institute of Cell Biology, University of Edinburgh, Edinburgh, EH9 3BF, UK.
Nuclei can be relatively easily extracted from homogenized liver due to the softness of the tissue and crudely separated from other cellular organelles by low-speed centrifugation due to the comparatively large size of nuclei. However, further purification is complicated by nuclear envelope continuity with the endoplasmic reticulum, invaginations containing mitochondria, and connections to the cytoskeleton. Subsequent purification to nuclear envelopes is additionally confounded by connections of inner nuclear membrane proteins to chromatin.
View Article and Find Full Text PDFOff-Line Multidimensional Liquid Chromatography and Auto Sampling Result in Sample Loss in LC/LC-MS/MS.
J Proteome Res
August 2014
†Department of Chemical Physiology, The Scripps Research Institute, 10550 North Torrey Pines Road, SR11, La Jolla, California 92037, United States.
Large-scale proteomics often employs two orthogonal separation methods to fractionate complex peptide mixtures. Fractionation can involve ion exchange separation coupled to reversed-phase separation or, more recently, two reversed-phase separations performed at different pH values. When multidimensional separations are combined with tandem mass spectrometry for protein identification, the strategy is often referred to as multidimensional protein identification technology (MudPIT).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!