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Proteomics-based method for the assessment of marine pollution using liquid chromatography coupled with two-dimensional electrophoresis
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
2007 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 6, no 6, 2094-2104 p.Article in journal (Refereed) Published
Abstract [en]

Using a proteomic approach, we have developed a new method for the assessment of marine pollution that generates highly reproducible protein expression patterns and it is simple and scalable. The protocol is based on applying liquid chromatography ( LC) coupled with two-dimensional electrophoresis (2DE) to analyze changes in the protein expression pattern after exposure to marine pollution. The digestive gland of the sentinel "blue mussel" ( Mytilus edulis) was batch-processed through a simple cell fractionation followed by ion-exchange chromatography and 2-DE. The selection of ligands, elution method, and small volume design was carefully considered to define a protocol that could be mainly robotized. A pilot study with samples collected from different Gothenburg harbor areas indicated that the clean area could be distinguished from the polluted ones based on a protein expression pattern ( PES) composed of 13 proteins. Principal component analysis ( PCA) and hierarchical clustering confirmed that the PES was sufficient to discriminate polluted and unpolluted areas and to provide a spatial gradient from the polluted source. Several proteins from the PES were identified by electrospray ionization tandem mass spectrometry ( ESI-MS/MS), and they are involved in,beta-oxidation, amino acid metabolism, detoxification, protein degradation, organelle biogenesis, and protein folding. In the near future, this methodology could show potential advantages to assess marine pollution and could become a stable platform to elucidate ecotoxicological questions.

Place, publisher, year, edition, pages
2007. Vol. 6, no 6, 2094-2104 p.
Keyword [en]
proteomics, biomarker profiling, liquid chromatography, two-dimensional electrophoresis, marine pollution assessment
National Category
Biological Sciences Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-145308DOI: 10.1021/pr060689sISI: 000246893500004OAI: oai:DiVA.org:uu-145308DiVA: diva2:395921
Available from: 2011-02-08 Created: 2011-02-08 Last updated: 2011-02-08Bibliographically approved

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