Artículos de revistas
Expression and purification of a small heat shock protein from the plant pathogen Xylella fastidiosa
Registro en:
Protein Expression And Purification. Academic Press Inc Elsevier Science, v. 33, n. 2, n. 297, n. 303, 2004.
1046-5928
WOS:000188414200017
10.1016/j.pep.2003.10.007
Autor
Azzoni, AR
Tada, SFS
Rosselli, LK
Paula, DP
Catani, CF
Sabino, AA
Barbosa, JARG
Guimaraes, BG
Eberlin, MN
Medrano, FJ
Souza, AP
Institución
Resumen
The small heat shock proteins (smHSPs) belong to a family of proteins that function as molecular chaperones by preventing protein aggregation and are also known to contain a conserved region termed a-crystallin domain. Here, we report the expression, purification, and partial characterization of a novel smHSP (HSP17.9) from the phytopathogen Xylella fastidiosa, causal agent of the citrus variegated chlorosis (CVC). The gene was cloned into a pET32-Xa/LIC vector to over-express the protein coupled with fusion tags in Escherichia coli BL21(DE3). The expressed HSP17.9 was purified by immobilized metal affinity chromatography (IMAC) and had its identity determined by mass spectrometry (MALDI-TOF). The correct folding of the purified recombinant protein was verified by circular dichroism spectroscopy. Finally, the HSP17.9 protein also proved to efficiently prevent induced aggregation of insulin, strongly indicating a chaperone-like activity. (C) 2003 Elsevier Inc. All rights reserved. 33 2 297 303