One-dimensional and two-dimensional polyacrylamide gel electrophoresis: a tool for protein characterisation in aquatic samples

Jones, Vera; Ruddell, Carolyn J.; Wainwright, Geoff; Rees, Huw H.; Jaffé, Rudolf; Wolff, George A.

doi:10.1016/j.marchem.2003.09.003

Cited by 20 publications

(8 citation statements)

References 31 publications

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“…Another way to assess the sources, cycling, and distribution of dissolved proteins is to target abundant proteins using 2D gel electrophoresis and sensitive immunoassays such as ELISA (enzyme-linked immunosorbent assay) and MSIA (mass spectrometry immunoassay; Jones et al, 2004;Orellana and Hansell, 2013;Orellana et al, 2003). In their proteomic data, reported short peptide sequences suggestive of ribulose-1,5-bisphosphate carboxylate/ oxygenase (RuBisCo).…”

Section: B Proteins and Amino Acids In Dommentioning

confidence: 99%

Chemical Characterization and Cycling of Dissolved Organic Matter

Repeta

2015

Biogeochemistry of Marine Dissolved Organic Matter

113

130

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Section: B Proteins and Amino Acids In Dommentioning

confidence: 99%

Chemical Characterization and Cycling of Dissolved Organic Matter

Repeta

2015

Biogeochemistry of Marine Dissolved Organic Matter

113

130

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“…Initial characterization studies involved quantitative analysis of amino acid residues through high temperature acid hydrolysis of water samples, followed by analysis using high performance liquid chromatography (HPLC) with ultraviolet or fluorescence detection (Lindroth and Mopper 1979). This approach has Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography-electrospray ionization-mass spectrometry Philippa yielded useful quantitative information, while analysis of the enantiomeric ratios of amino acids can be used to track sources (Jones et al 2005). However, these approaches provide little information on molecular structure, and some of the amino acid residues have low recoveries (Keil and Kirchman 1991;Tappin et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography‐electrospray ionization‐mass spectrometry

Curtis-Jackson

Massé

Gledhill

et al. 2009

Limnology & Ocean Methods

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Low molecular weight (LMW; <1 kDa) dissolved organic nitrogen (DON) is the dominant DON fraction in marine systems. However, existing methods for DON characterization exclude this fraction through size exclusion or destruction during sample hydrolysis. In this study, we developed a method for the extraction and analysis of LMW peptides in saline waters. The procedure involves a solid phase extraction (SPE) preconcentration step using a polystyrene-divinylbenzene sorbent, then elution and detection by liquid chromatographyelectrospray ionization-mass spectrometry (LC/ESI/MS). The procedure was tested with 1-L water samples containing peptides (10 ng L -1 ) ranging from 188.2-1946.0 Da. The analytes were characterized using "top-down" sequencing to confirm their structure, and the method was then successfully applied to saline water samples extracted from a mono-algal culture of Phaeodactylum tricornutum. This enabled a number of unknown analytes to be partially sequenced (complete sequencing should be possible using additional MS n scans and larger sample volumes). The method allows for the detection of peptides at the ng L -1 level and further preconcentration is possible. The SPE step allowed determination of peptides from saline water, a matrix incompatible with direct LC/MS analysis. The method can, therefore, be used in DON characterization studies.

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“…Two-dimensional electrophoresis was performed according to methods (Sanchez et al, 1997;Cornillot et al, 2002;Felicio et al, 2003;Jones et al, 2004) and the operational manual of Amersham Pharmacia Biotech, Sweden. It mainly included the following: 1) Isoelectric focusing (IEF) of proteins on pre-prepared immobilized gradient (IPG) strip.…”

Section: D-pagementioning

confidence: 99%

Two-dimensional electrophoresis analysis of proteins extracted from Alexandrium sp. LC3

Miao²,

et al. 2007

J Ocean Univ. China

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Two-dimensional electrophoresis(2-DE) of protein extracted and purified from Alexandrium sp. LC3 was conducted. In the SDS-PAGE study, the relative molecular weights of the proteins were mainly in the range of 14 kDa−31 kDa and 43 kDa−66 kDa, and more proteins were detected between 14 kDa and 31 kDa. With the improved protein preparation, the two-dimensional electrophoresis patterns indicated that the relative molecular weights of the proteins were between 14 kDa and 100 kDa, and most of them ranged from 14 kDa to 31 kDa. This was consistent with the result of the SDS-PAGE analysis. The isoelectric points were found to lie between 3.0 and 8.0, and most of them were in the range of 3.0−6.0. Better separation effect was acquired with pre-prepared immobilized gradient (IPG) strip (pH 3−5.6), and about 320 protein spots could be visualized on the 2-DE map by staining. Within pH 3−10 and pH 3−5.6 strips, the protein samples of Alexandrium sp. LC3 could be separated well.

show abstract

One-dimensional and two-dimensional polyacrylamide gel electrophoresis: a tool for protein characterisation in aquatic samples

Cited by 20 publications

References 31 publications

Chemical Characterization and Cycling of Dissolved Organic Matter

Chemical Characterization and Cycling of Dissolved Organic Matter

Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography‐electrospray ionization‐mass spectrometry

Two-dimensional electrophoresis analysis of proteins extracted from Alexandrium sp. LC3

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