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Development and application of a proteomic approach to the assessment of pollution in the marine environment
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics. (Environmental Proteomics)
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Today, assessment of the health of coastal waters is recognized as being important for both the conservation of nature and well-being of humans. Anthropogenic pollution has been the focus of extensive research for some time and a variety of programs for the monitoring and assessment of environmental pollution have been developed. Determination of the levels of pollution in sensitive ‘sentinels’ such as mussels, allows monitoring of these levels in a given area over a prolonged period of time. Furthermore, the biological effects of pollution are reflected in a series of biomarkers, none of which provides a general picture of the sentinel’s state of health and all of which are individually specific for certain pollutants and influenced by both biotic and abiotic factors.

In an attempt to improve biomonitoring of marine pollution, we have developed two proteomic approaches here. In the first portion of the thesis, a proteomic analysis was performed on peroxisomes isolated from mussels exposed either to one of three model anthropogenic pollutants, or two different types of crude oil, or from mussels exposed to the Prestige oil spill. Application of two-dimensional electrophoresis (2-DE) provided protein expression signatures (PES) for exposure to these different pollutants.Furthermore, several individual protein components of these PES could be putatively identified.

In the second portion of this work, such analysis of subproteomes was developed further in order to improve the applicability of this approach to biomonitoring. A simple fractionation procedure in combination with liquid chromatography and 2-DE provided samples from mussels residing in different regions of a pollution gradient around the harbor of Gothenburg, as well as from mussels exposed to two types of fuel oil similar to that of the Prestige that were suitable for environmental proteomics. In addition, we constructed a model for this approach that can be cross-validated in the future and applied to assess sources of fuel oil pollution in connection with biomonitoring programs.

Place, publisher, year, edition, pages
Stockholm: Department of Biochemistry and Biophysics, Stockholm University , 2009. , 58 p.
Keyword [en]
proteomics, mussels, peroxisomes, two-dimensional electrophoresis (2-DE), difference gel electrophoresis (DIGE), bisphenol A (BPA), diallyl phthalate (DAP), 2, 2′, 4, 4′-tetrabromodiphenyl ether (PBDE-47), fuel oil, Prestige oil spill
National Category
Environmental Sciences Pharmacology and Toxicology
Research subject
Biochemistry
Identifiers
URN: urn:nbn:se:su:diva-26150ISBN: 978-91-7155-801-5 (print)OAI: oai:DiVA.org:su-26150DiVA: diva2:207352
Public defence
2009-04-17, Magnélisalen , Kemiska övningslaboratoriet, Svante Arrhenius väg 12A, Stockholm University, Stockholm, 14:00 (English)
Opponent
Supervisors
Available from: 2009-03-26 Created: 2009-03-10 Last updated: 2009-04-02Bibliographically approved
List of papers
1. Identification of proteomic signatures of exposure to marine pollutants in mussels (Mytilus edulis)
Open this publication in new window or tab >>Identification of proteomic signatures of exposure to marine pollutants in mussels (Mytilus edulis)
2006 (English)In: Molecular & Cellular Proteomics, ISSN 1535-9476, Vol. 5, no 7, 1274-1285 p.Article in journal (Refereed) Published
Keyword
Animals, Electrophoresis; Gel; Two-Dimensional, Environmental Exposure, Gene Expression Profiling, Multivariate Analysis, Mytilus edulis/*drug effects/*metabolism, Proteomics, Water Pollutants/*adverse effects
Identifiers
urn:nbn:se:su:diva-11254 (URN)10.1074/mcp.M500333-MCP200 (DOI)
Available from: 2008-01-10 Created: 2008-01-10 Last updated: 2009-03-24Bibliographically approved
2. Peroxisomal proteomic approach for protein profiling in blue mussels (Mytilus edulis) exposed to crude oil
Open this publication in new window or tab >>Peroxisomal proteomic approach for protein profiling in blue mussels (Mytilus edulis) exposed to crude oil
2007 (English)In: Biomarkers, ISSN 1354-750X, Vol. 12, no 1, 47-60 p.Article in journal (Refereed) Published
Keyword
Animals, Bivalvia/*chemistry, Electrophoresis; Gel; Two-Dimensional, Peroxisomes/chemistry/*drug effects, Petroleum/*toxicity, Proteins/*analysis, Proteomics, Water Pollutants; Chemical/*toxicity
Identifiers
urn:nbn:se:su:diva-11253 (URN)10.1080/13547500600943528 (DOI)000243660200004 ()
Available from: 2008-01-10 Created: 2008-01-10 Last updated: 2009-03-24Bibliographically approved
3. Peroxisomal proteomics, biomonitoring in mussles after the Prestige's oil spill
Open this publication in new window or tab >>Peroxisomal proteomics, biomonitoring in mussles after the Prestige's oil spill
2009 (English)In: Marine Pollution Bulletin, ISSN 0025-326X, E-ISSN 1879-3363, Vol. 58, no 12, 1815-1826 p.Article in journal (Refereed) Published
Abstract [en]

Peroxisomal proteomics was applied to assess possible biological effects after the Prestige's oil spill. Mussels were sampled in July 2004 and 2005 in four stations in the NW (closest to the spill) and NE coasts of the Iberian Peninsula. Principal components analysis (PCA) suggested differences in protein expression among stations and sampling years. Several proteins were putatively identified by mass spectrometry and immunolocalization. PC1 separated the NW stations in 2004 from the rest of the stations and sampling years mainly due to up-regulation of peroxisomal β-oxidation proteins and PMP70. PC3 separated the NE-stations, based on up-regulation of the antioxidant enzyme catalase in 2004 compared to 2005. PC4 separated the stations in the NE and the NW. This work shows that environmental proteomics, together with multivariate data analysis, could provide information to interpret the effects of oil spills at cellular level in mussels also in the absence of historical data.

Keyword
proteomics, biomarker, two-dimensional electrophoresis, marine pollution assessment, oil spill, Prestige
Identifiers
urn:nbn:se:su:diva-26585 (URN)10.1016/j.marpolbul.2009.08.004 (DOI)000273104200018 ()
Available from: 2009-04-02 Created: 2009-04-02 Last updated: 2010-08-11Bibliographically approved
4. Proteomics-based method for the assessment of marine pollution using liquid chromatography coupled with two-dimensional electrophoresis
Open this publication in new window or tab >>Proteomics-based method for the assessment of marine pollution using liquid chromatography coupled with two-dimensional electrophoresis
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 (2-DE) 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 β-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.

Keyword
Amino Acid Sequence, Animals, Cell Fractionation, Chromatography; Ion Exchange/methods, Electrophoresis; Gel; Two-Dimensional/methods, Molecular Sequence Data, Mytilus edulis/*chemistry, Oceans and Seas, Proteins/*analysis, Proteomics/*methods, Spectrometry; Mass; Electrospray Ionization, Water Pollution; Chemical
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:su:diva-11250 (URN)10.1021/pr060689s (DOI)000246893500004 ()
Available from: 2008-01-10 Created: 2008-01-10 Last updated: 2011-04-20Bibliographically approved
5. Proteomic analysis of mussels exposed to fresh and weathered Prestige's oil
Open this publication in new window or tab >>Proteomic analysis of mussels exposed to fresh and weathered Prestige's oil
Show others...
(English)Manuscript (Other academic)
Abstract [en]

Biomonitoring
programs
that
use
mussels to assess the
water
quality
around
the
world, could
benefit
from
the
use
 of
proteomics
techniques. These
could
be
applied to obtain
protein expression
signatures
of
exposure to
pollution
that
could
be further
used
for
prediction purposes. This
would
require
that
a
combination
of
univariate
and
multivariate
 statistical
analyses
of
proteomics data
were
utilized
to
obtain
robust
models.
We
show an
application
of
this approach
on
mussels
exposed
to
fresh
fuel, and weathered
fuel
in
a
laboratory experiment
that
tried
to
mimic
the
effects of the
Prestige's
oil
spill.
A
set
of
protein
spots
were
selected
that
could
be used
to
classify
mussels exposed
to
the two
types
of
fuel
oil.
As
an
example
of the possibilities
this
approach
could offer to biomonitoring
programs, mussels were collected from ten sampling stations
along
the
NW
and
NE of the Iberian Peninsula, and their protein expression patterns monitored.

 

 

Keyword
Prestige oil spill, fresh fuel, weathered fuel, mussels, two-dimensional electrophoresis (2-DE), two-way ANOVA, false discovery rate (FDR), principal components analysis (PCA)
Identifiers
urn:nbn:se:su:diva-26391 (URN)
Available from: 2009-03-23 Created: 2009-03-23 Last updated: 2010-01-14Bibliographically approved

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