Evaluation of protein extraction methods for enhanced proteomic analysis of tomato leaves and roots

Autor(es)

Vilhena, Milca B. ^[1] ; Franco, Monica R. ^[1] ; Schmidt, Daiana ^[1] ; Carvalho, Giselle ^[1] ; Azevedo, Ricardo A. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Genet 11, ESALQ, BR-13400970 Piracicaba, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Anais da Academia Brasileira de Ciências; v. 87, n. 3, p. 1853-1863, SEP 2015.
• Citações Web of Science: 6

Resumo

Proteomics is an outstanding area in science whose increasing application has advanced to distinct purposes. A crucial aspect to achieve a good proteome resolution is the establishment of a methodology that results in the best quality and wide range representation of total proteins. Another important aspect is that in many studies, limited amounts of tissue and total protein in the tissue to be studied are found, making difficult the analysis. In order to test different parameters, combinations using minimum amount of tissue with 4 protocols for protein extraction from tomato (Solanum lycopersicum L.) leaves and roots were evaluated with special attention to their capacity for removing interferents and achieving suitable resolution in bidimensional gel electrophoresis, as well as satisfactory protein yield. Evaluation of the extraction protocols revealed large protein yield differences obtained for each one. TCA/acetone was shown to be the most efficient protocol, which allowed detection of 211 spots for leaves and 336 for roots using 500 µg of leaf protein and 800 µg of root protein per gel. (AU)

• Processo FAPESP: 09/54676-0 - Estresse oxidativo induzido por metais: novas abordagens
• Beneficiário: Ricardo Antunes de Azevedo
• Modalidade de apoio: Auxílio à Pesquisa - Temático