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Overexpression, purification, crystallization and preliminary X-ray diffraction analysis of the F1 antigen Caf1M-Caf1 chaperone-subunit pre-assembly complex from Yersinia pestis.

Overexpression, purification, crystallization and preliminary X-ray diffraction analysis of the F1 antigen Caf1M-Caf1 chaperone-subunit pre-assembly complex from Yersinia pestis.

Authors :
Zavialov A
Berglund J
Knight SD
Source :
Acta crystallographica. Section D, Biological crystallography [Acta Crystallogr D Biol Crystallogr] 2003 Feb; Vol. 59 (Pt 2), pp. 359-62. Date of Electronic Publication: 2003 Jan 23.
Publication Year :
2003

Abstract

The F1 capsular antigen of the plague-causing pathogen Yersinia pestis is assembled from monomeric Caf1 subunits via the Caf1M/Caf1A chaperone/usher system. Y. pestis Caf1M-Caf1 chaperone-subunit complex was purified from the periplasm of Escherichia coli cells overexpressing Caf1M and Caf1 and was crystallized in PEG 4000 solution using hanging-drop vapour diffusion. The crystals diffract to a minimum Bragg spacing of 1.8 A and belong to space group P2(1), with unit-cell parameters a = 36.0, b = 69.2, c = 69.1 A, beta = 93.0 degrees. SeMet-labelled Caf1M-Caf1 complexes were purified and crystallized under the same conditions. The SeMet crystals were identical to the native crystals and diffracted to 1.9 A. Heavy-atom derivative crystals were prepared by soaking in 10 mM K(2)PtCl(4), giving two Pt sites per complex. The experimental electron-density map was obtained by a combination of MAD and MIR methods using both Se- and Pt-derivative crystals.

Details

Language :
English
ISSN :
0907-4449
Volume :
59
Issue :
Pt 2
Database :
MEDLINE
Journal :
Acta crystallographica. Section D, Biological crystallography
Publication Type :
Academic Journal
Accession number :
12554953
Full Text :
https://doi.org/10.1107/s0907444902021054