A novel method for phosphoprotein extraction from macroalgae
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María L. Parages
Abstract
The extraction of proteins from macroalgae has always been difficult, primarily because of their high internal levels of polysaccharides and polyphenols. Several methods for protein extraction have been developed recently that allow the production of high quality extracts for 1-D SDS gel electrophoresis or 2-D SDS-polyacrylamide gel electrophoresis for proteomic analysis. However, none of the existing methods are applicable to the study of the phosphorylation state of kinases involved in stress responses of macroalgae, primarily because trichloroacetic acid and phenol are used for protein precipitation. In this work, we describe a novel method for protein extraction from macroalgae based on the counterintuitive use of commercial RNA purification kits; this method maintains the phosphorylation state of the kinases intact, as demonstrated by either polyclonal or monoclonal antibodies against the phosphorylated forms of mitogen-activated protein kinases using Western blot methodology. This method may be appropriate for wider use in the analysis of phosphoproteomes.
©2012 by Walter de Gruyter Berlin Boston
Artikel in diesem Heft
- Masthead
- Masthead
- Research Articles
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- New record of Sargassum denticarpum Ajisaka (Fucales, Phaeophyceae) from Iriomote Island (Ryukyu Archipelago, Japan)
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- Morphological and molecular evidence demonstrates the amphi-Atlantic distribution of Laurencia catarinensis (Ceramiales, Rhodophyta)
- Effects of temperature, salinity, irradiance, and nutrients on the development of carposporelings and tetrasporophytes in Gracilaria domingensis (Kütz.) Sonder ex Dickie (Rhodophyta, Gracilariales)
- A novel method for phosphoprotein extraction from macroalgae
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- The marine halophytes Carpobrotus edulis L. and Arthrocnemum macrostachyum L. are potential sources of nutritionally important PUFAs and metabolites with antioxidant, metal chelating and anticholinesterase inhibitory activities
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- Short Communications
- Application of image analysis software to the morphometrics of Posidonia oceanica (L.) Delile and Cymodocea nodosa (Ucria) Ascherson
- Delayed modifications in plant-water relations in the coastal marsh halophyte Spartina patens following sudden increases in soil salinity
- Algicidal activity and potential antifouling defenses in macroalgae from the western Antarctic Peninsula including probable synergistic effects of multiple compounds
Artikel in diesem Heft
- Masthead
- Masthead
- Research Articles
- Fucoid assemblages on the north coast of Spain: past and present (1977–2007)
- New record of Sargassum denticarpum Ajisaka (Fucales, Phaeophyceae) from Iriomote Island (Ryukyu Archipelago, Japan)
- Seasonal periodicity and reproduction of brown algae (Phaeophyceae) at Puerto Deseado (Patagonia)
- Determining bathymetric distributions of the eelgrass Zostera marina L. in three turbid estuaries on the eastern North Pacific coast
- Morphological and molecular evidence demonstrates the amphi-Atlantic distribution of Laurencia catarinensis (Ceramiales, Rhodophyta)
- Effects of temperature, salinity, irradiance, and nutrients on the development of carposporelings and tetrasporophytes in Gracilaria domingensis (Kütz.) Sonder ex Dickie (Rhodophyta, Gracilariales)
- A novel method for phosphoprotein extraction from macroalgae
- Antioxidants in mangrove plants and endophytic fungal associations
- The marine halophytes Carpobrotus edulis L. and Arthrocnemum macrostachyum L. are potential sources of nutritionally important PUFAs and metabolites with antioxidant, metal chelating and anticholinesterase inhibitory activities
- Culture medium optimization and lipid profiling of Cylindrotheca, a lipid- and polyunsaturated fatty acid-rich pennate diatom and potential source of eicosapentaenoic acid
- Short Communications
- Application of image analysis software to the morphometrics of Posidonia oceanica (L.) Delile and Cymodocea nodosa (Ucria) Ascherson
- Delayed modifications in plant-water relations in the coastal marsh halophyte Spartina patens following sudden increases in soil salinity
- Algicidal activity and potential antifouling defenses in macroalgae from the western Antarctic Peninsula including probable synergistic effects of multiple compounds
