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Four different detergents, ASB 14, SB 3-10, CHAPS and Triton X100, were utilized to determine the optimal detergent for the solubilization of membrane proteins from the phytopathogenic bacterium Xylella fastidiosa. These proteins were differentially solubilized in distinct buffers containing the detergent and subjected to bidimensional electrophoresis within the non-linear pH range of 3-10. The detergents ASB 14 and SB 3-10 were the most effective revealing 221 and 157 spots, respectively. CHAPS and Triton X100 were less effective and revealed only 72 and 43 spots, respectively. MALDI-TOF tryptic peptide mass fingerprinting of 18 excised proteins from the ASB 14 treatment revealed that 83% were membrane proteins and that the theoretical efficiency of solubilization for ASB 14 was estimated to be 87%. This study demonstrates the effectiveness of the detergent ASB 14 for the solubilization of membrane proteins from the bacterium X. fastidiosa. O objetivo deste trabalho foi comparar a eficiência da solubilização de quatro detergentes comercialmente disponíveis, ASB 14, SB 3-10, CHAPS e Triton X100, na extração de proteínas de membrana da bactéria Xylella fastidiosa para estudos proteômicos. Estas proteínas foram solubilizadas em duas etapas em tampões diferenciados pelos detergentes e submetidas à eletroforese bidimensional (2-DE) em uma faixa de pH não linear de 3-10. Os detergentes ASB 14 e SB 3-10 foram os mais efecientes, revelando 221 e 157 proteínas, respectivamente, enquanto que o CHAPS e o Triton X100 resultaram somente 72 e 43 proteínas, respectivamente. A identificação das proteínas foi feita por 'peptide mass fingerprinting' em espectrometria de massa MALDI-TOF, através de peptídeos obtidos por digestão com tripsina in gel. Os 18 spots de proteínas do gel com tratamento com ASB 14 mostrou que 83% eram proteínas de membrana. Este estudo concluiu que o detergente ASB-14 foi o mais eficiente na solubilização de proteínas de membrana de Xylella fastidiosa

Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis