Method for obtaining high-resolution proteomic analysis from pericarps of guarana
Guarana has great agricultural potential and is largely used therapeutically and in the production of non-alcoholic energy drinks. Genomic and proteomic studies are crucial to identify proteins that play central roles in the maintenance and viability of fruits, as well as to identify proteins relate...
| Autor principal: | Souza, Andréa L. |
|---|---|
| Outros Autores: | Angelo, P., Nogueira, P. P O, Gonçalves, José Francisco Carvalho de, Franco, A. M., Astolfi-Filho, Spártaco A.T., López-Lozano, Jorge Luis, Andrade, Edmar V. |
| Grau: | Artigo |
| Idioma: | English |
| Publicado em: |
Genetics and Molecular Research
2020
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| Assuntos: | |
| Acesso em linha: |
https://repositorio.inpa.gov.br/handle/1/14987 |
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oai:repositorio:1-14987 |
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dspace |
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oai:repositorio:1-14987 Method for obtaining high-resolution proteomic analysis from pericarps of guarana Souza, Andréa L. Angelo, P. Nogueira, P. P O Gonçalves, José Francisco Carvalho de Franco, A. M. Astolfi-Filho, Spártaco A.T. López-Lozano, Jorge Luis Andrade, Edmar V. Acetone Ascorbate Peroxidase Carbohydrate Cyclophilin Sodium Dodecyl Sulfate Enolase Glyceraldehyde 3 Phosphate Dehydrogenase Histone H2b Lectin Lipid Phenol Povidone Superoxide Dismutase Trichloroacetic Acid Vegetable Protein Contamination Extraction Guarana Nonhuman Pericarp Precipitation Protein Analysis Proteomics Solubility Tandem Mass Spectrometry Two Dimensional Electrophoresis Chemistry Electrospray Mass Spectrometry Metabolism Paullinia Two Dimensional Gel Electrophoresis Paullinia Cupana Electrophoresis, Gel, Two-dimensional Paullinia Plant Proteins Proteomics Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry Guarana has great agricultural potential and is largely used therapeutically and in the production of non-alcoholic energy drinks. Genomic and proteomic studies are crucial to identify proteins that play central roles in the maintenance and viability of fruits, as well as to identify proteins related to the main metabolic pathways. However, the success of any protein analysis starts with the protein extract preparation, which needs to offer an extract that is free of contaminants. This study aimed to evaluate different extraction methods to obtain high-quantity and high-quality extracts that are compatible with analysis by 2-dimensional electrophoresis and tandem mass spectrometry protein identification. Three different methods were tested: trichloroacetic acid (TCA)/acetone, sodium dodecyl sulfate (SDS)/phenol, and polyvinylpolypyrrolidone (PVPP)/SDS/phenol. The extract obtained from the TCA/acetone precipitation presented low solubility and contamination with lipids and carbohydrates. On the other hand, the quality of the extract gradually improved after using phenol and PVPP/phenol, enabling a yield up to 2 mg/g macerated tissues and the detection of 457 spots by 2-dimensional electrophoresis. The effectiveness of the procedure used was validated by identification of 10 randomly selected proteins by mass spectrometry. The procedure described here can be a starting point for applications using tissues of other organs of guarana or tissues of species that are similar to guarana. © FUNPEC-RP. 2020-05-07T14:00:21Z 2020-05-07T14:00:21Z 2014 Artigo https://repositorio.inpa.gov.br/handle/1/14987 10.4238/2014.September.29.14 en Volume 13, Número 3, Pags. 8014-8024 Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ application/pdf Genetics and Molecular Research |
| institution |
Instituto Nacional de Pesquisas da Amazônia - Repositório Institucional |
| collection |
INPA-RI |
| language |
English |
| topic |
Acetone Ascorbate Peroxidase Carbohydrate Cyclophilin Sodium Dodecyl Sulfate Enolase Glyceraldehyde 3 Phosphate Dehydrogenase Histone H2b Lectin Lipid Phenol Povidone Superoxide Dismutase Trichloroacetic Acid Vegetable Protein Contamination Extraction Guarana Nonhuman Pericarp Precipitation Protein Analysis Proteomics Solubility Tandem Mass Spectrometry Two Dimensional Electrophoresis Chemistry Electrospray Mass Spectrometry Metabolism Paullinia Two Dimensional Gel Electrophoresis Paullinia Cupana Electrophoresis, Gel, Two-dimensional Paullinia Plant Proteins Proteomics Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry |
| spellingShingle |
Acetone Ascorbate Peroxidase Carbohydrate Cyclophilin Sodium Dodecyl Sulfate Enolase Glyceraldehyde 3 Phosphate Dehydrogenase Histone H2b Lectin Lipid Phenol Povidone Superoxide Dismutase Trichloroacetic Acid Vegetable Protein Contamination Extraction Guarana Nonhuman Pericarp Precipitation Protein Analysis Proteomics Solubility Tandem Mass Spectrometry Two Dimensional Electrophoresis Chemistry Electrospray Mass Spectrometry Metabolism Paullinia Two Dimensional Gel Electrophoresis Paullinia Cupana Electrophoresis, Gel, Two-dimensional Paullinia Plant Proteins Proteomics Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry Souza, Andréa L. Method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| topic_facet |
Acetone Ascorbate Peroxidase Carbohydrate Cyclophilin Sodium Dodecyl Sulfate Enolase Glyceraldehyde 3 Phosphate Dehydrogenase Histone H2b Lectin Lipid Phenol Povidone Superoxide Dismutase Trichloroacetic Acid Vegetable Protein Contamination Extraction Guarana Nonhuman Pericarp Precipitation Protein Analysis Proteomics Solubility Tandem Mass Spectrometry Two Dimensional Electrophoresis Chemistry Electrospray Mass Spectrometry Metabolism Paullinia Two Dimensional Gel Electrophoresis Paullinia Cupana Electrophoresis, Gel, Two-dimensional Paullinia Plant Proteins Proteomics Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry |
| description |
Guarana has great agricultural potential and is largely used therapeutically and in the production of non-alcoholic energy drinks. Genomic and proteomic studies are crucial to identify proteins that play central roles in the maintenance and viability of fruits, as well as to identify proteins related to the main metabolic pathways. However, the success of any protein analysis starts with the protein extract preparation, which needs to offer an extract that is free of contaminants. This study aimed to evaluate different extraction methods to obtain high-quantity and high-quality extracts that are compatible with analysis by 2-dimensional electrophoresis and tandem mass spectrometry protein identification. Three different methods were tested: trichloroacetic acid (TCA)/acetone, sodium dodecyl sulfate (SDS)/phenol, and polyvinylpolypyrrolidone (PVPP)/SDS/phenol. The extract obtained from the TCA/acetone precipitation presented low solubility and contamination with lipids and carbohydrates. On the other hand, the quality of the extract gradually improved after using phenol and PVPP/phenol, enabling a yield up to 2 mg/g macerated tissues and the detection of 457 spots by 2-dimensional electrophoresis. The effectiveness of the procedure used was validated by identification of 10 randomly selected proteins by mass spectrometry. The procedure described here can be a starting point for applications using tissues of other organs of guarana or tissues of species that are similar to guarana. © FUNPEC-RP. |
| format |
Artigo |
| author |
Souza, Andréa L. |
| author2 |
Angelo, P. Nogueira, P. P O Gonçalves, José Francisco Carvalho de Franco, A. M. Astolfi-Filho, Spártaco A.T. López-Lozano, Jorge Luis Andrade, Edmar V. |
| author2Str |
Angelo, P. Nogueira, P. P O Gonçalves, José Francisco Carvalho de Franco, A. M. Astolfi-Filho, Spártaco A.T. López-Lozano, Jorge Luis Andrade, Edmar V. |
| title |
Method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| title_short |
Method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| title_full |
Method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| title_fullStr |
Method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| title_full_unstemmed |
Method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| title_sort |
method for obtaining high-resolution proteomic analysis from pericarps of guarana |
| publisher |
Genetics and Molecular Research |
| publishDate |
2020 |
| url |
https://repositorio.inpa.gov.br/handle/1/14987 |
| _version_ |
1787144534649470976 |
| score |
11.755432 |