Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins

Kirkland, P. Aaron; Busby, Jennifer; Stevens, Stanley M.; Maupin-Furlow, Julie A.

doi:10.1016/j.ab.2006.01.017

Cited by 63 publications

(77 citation statements)

References 10 publications

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“…Total protein extraction was performed based on previous studies with some modifications [16,17]. Briefly, the mussel samples were homogenized quickly on ice with 1 mL of TRIzol reagent and centrifuged at 12000 g for 5 min at 4 C. The supernatant was added with 200 mL of chloroform before shaking vigorously for 3 min and precipitating for 3 min.…”

Section: Protein Extraction Of Mussel Samplesmentioning

confidence: 99%

Proteomic responses reveal the differential effects induced by cadmium in mussels Mytilus galloprovincialis at early life stages

Peng

et al. 2016

Fish & Shellfish Immunology

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a b s t r a c tCadmium (Cd) has become an important metal contaminant and posed severe risk on the organisms in the coastal environments of the Bohai Sea. Marine mussel Mytilus galloprovincialis is widely distributed along the Bohai coast and consumed as seafood by local residents. Evidences indicate that the early stages of marine organisms are more sensitive to metal contaminants. In this study, we applied twodimensional electrophoresis-based proteomics to characterize the biological effects of Cd (50 mg L À1 ) in the early life stages (D-shape larval and juvenile) of mussels. The different proteomic responses demonstrated the differential responsive mechanisms to Cd exposure in these two early life stages of mussels. In details, results indicated that Cd mainly induced immune and oxidative stresses in both Dshape larval and juvenile mussels via different pathways. In addition, the significant up-regulation of triosephosphate isomerase and metallothionein confirmed the enhanced energy demand and mobilized detoxification mechanism in D-shape larval mussels exposed to Cd. In juvenile mussels, Cd exposure also induced clear apoptosis. Overall, this work suggests that Cd is a potential immune toxicant to mussel M. galloprovincialis at early life stages.

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Section: Protein Extraction Of Mussel Samplesmentioning

confidence: 99%

Proteomic responses reveal the differential effects induced by cadmium in mussels Mytilus galloprovincialis at early life stages

Peng

et al. 2016

Fish & Shellfish Immunology

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“…Total protein extraction was applied based on Lee et al [26] and Kirkland et al [27] with some modifications. Briefly, the tissue of hepatopancreas was homogenized quickly on ice with 1 mL of Trizol reagent and centrifuged at 12,000 g for 5 min at 4°C.…”

Section: Protein Extractionmentioning

confidence: 99%

Proteomic and metabolomic responses in hepatopancreas of Mytilus galloprovincialis challenged by Micrococcus luteus and Vibrio anguillarum

Wei

et al. 2013

Journal of Proteomics

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The outbreak of pathogens can induce diseases and lead to massive mortalities of aquaculture animals including fish, mollusk and shrimp. In this work, the responses induced by Micrococcus luteus and Vibrio anguillarum were investigated in hepatopancreas of mussel Mytilus galloprovincialis using proteomics and metabolomics. Metabolic biomarkers demonstrated that M. luteus and V. anguillarum injections could induce osmotic stress and disturbance in energy metabolism. And the uniquely and more markedly altered metabolic biomarkers (glutamine, succinate, aspartate, glucose, ATP, homarine and tyrosine) indicated that V. anguillarum could cause more severe disturbances in osmotic regulation and energy metabolism. The differentially altered proteins meant that M. luteus and V. anguillarum induced different effects in mussels. However, the common proteomic biomarkers, arginine kinase and small heat shock protein, demonstrated that these two bacteria induced similar effects including oxidative stress and disturbance in energy metabolism in M. galloprovincialis. In addition, some metabolic biomarkers, ATP and glutamine, were confirmed by related proteins including arginine kinase, ATP synthase, nucleoside diphosphate kinase and glutamine synthetase in bacteria-challenged mussels.This study demonstrated that proteomics and metabolomics could provide an insightful view into the effects of environmental pathogens to the marine mussel M. galloprovincialis. Biological significanceThe outbreak of pathogens can lead to diseases and massive mortalities of aquaculture animals including fish, mollusk and shrimp. The mussel M. galloprovincialis distributes widely along the Bohai coast and is popularly consumed as delicious seafood by local residents. This bivalve has become one of the important species in marine aquaculture industry in China. Therefore a study on pathogen-induced effects is necessary. In the present study, an integrated metabolomic and proteomic approach was used to elucidate the differential effects induced by the representative Gram-positive (M. luteus) and Gram-negative (V. anguillarum) bacteria in M. galloprovincialis.

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“…Total protein extraction was applied based on the published method with some modifications [19,20]. Briefly, the gill tissue was homogenized quickly on ice with 1 ml Trizol reagent and centrifuged at 12 000 g for 5 min at 4 C. The supernatant was added with 200 mL chloroform before shaking vigorously for 3 min and precipitating for 3 min.…”

Section: Protein Extractionmentioning

confidence: 99%

Responses of Mytilus galloprovincialis to bacterial challenges by metabolomics and proteomics

Wei

et al. 2013

Fish & Shellfish Immunology

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a b s t r a c tPathogens can cause diseases and lead to massive mortalities of aquaculture animals and substantial economic loss. In this work, we studied the responses induced by Micrococcus luteus and Vibrio anguillarum in gill of mussel Mytilus galloprovincialis at protein and metabolite levels. Metabolic biomarkers (e.g., amino acids, betaine, ATP) suggested that both M. luteus and V. anguillarum induced disturbances in energy metabolism and osmotic regulation. The unique and some more remarkably altered metabolic biomarkers (threonine, alanine, aspartate, taurine, succinate) demonstrated that V. anguillarum could cause more severe disturbances in osmotic regulation and energy metabolism. Proteomic biomarkers (e.g., goose-type lysozyme 2, matrilin, ependymin-related protein, peptidylprolyl cisetrans isomerases) indicated that M. luteus caused immune stress, and disturbances in signaling pathways and protein synthesis. However, V. anguillarum mainly induced oxidative stress and disturbance in energy metabolism in mussel gills indicated by altered procollagen-proline dioxygenase, protein disulfide isomerase, nucleoside diphosphate kinases, electron transfer flavoprotein and glutathione S-transferase. This work confirmed that an integration of proteomics and metabolomics could provide an insightful view into the effects of pathogens to the marine mussel M. galloprovincialis.

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Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins

Cited by 63 publications

References 10 publications

Proteomic responses reveal the differential effects induced by cadmium in mussels Mytilus galloprovincialis at early life stages

Proteomic responses reveal the differential effects induced by cadmium in mussels Mytilus galloprovincialis at early life stages

Proteomic and metabolomic responses in hepatopancreas of Mytilus galloprovincialis challenged by Micrococcus luteus and Vibrio anguillarum

Responses of Mytilus galloprovincialis to bacterial challenges by metabolomics and proteomics

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