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

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.

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Main Authors: Di Ciero,Luciana, Bellato,Cláudia de M., Meinhardt,Lyndel W., Ferrari,Fernanda, Castellari,Rafael R., Marangoni,Sérgio, Novello,José C.
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Microbiologia 2004
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200018
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spelling oai:scielo:S1517-838220040002000182005-11-25Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresisDi Ciero,LucianaBellato,Cláudia de M.Meinhardt,Lyndel W.Ferrari,FernandaCastellari,Rafael R.Marangoni,SérgioNovello,José C. X. fastidiosa 2-DE extraction proteome solubilization 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.info:eu-repo/semantics/openAccessSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology v.35 n.3 20042004-09-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200018en10.1590/S1517-83822004000200018
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libraryname SciELO
language English
format Digital
author Di Ciero,Luciana
Bellato,Cláudia de M.
Meinhardt,Lyndel W.
Ferrari,Fernanda
Castellari,Rafael R.
Marangoni,Sérgio
Novello,José C.
spellingShingle Di Ciero,Luciana
Bellato,Cláudia de M.
Meinhardt,Lyndel W.
Ferrari,Fernanda
Castellari,Rafael R.
Marangoni,Sérgio
Novello,José C.
Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
author_facet Di Ciero,Luciana
Bellato,Cláudia de M.
Meinhardt,Lyndel W.
Ferrari,Fernanda
Castellari,Rafael R.
Marangoni,Sérgio
Novello,José C.
author_sort Di Ciero,Luciana
title Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
title_short Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
title_full Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
title_fullStr Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
title_full_unstemmed Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis
title_sort assessment of four different detergents used to extract membrane proteins from xylella fastidiosa by two-dimensional electrophoresis
description 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.
publisher Sociedade Brasileira de Microbiologia
publishDate 2004
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200018
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