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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.
- 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.
- Subjects :
- Antigens, Bacterial biosynthesis
Antigens, Bacterial genetics
Antigens, Bacterial isolation & purification
Bacterial Proteins biosynthesis
Bacterial Proteins genetics
Bacterial Proteins isolation & purification
Crystallization methods
Crystallography, X-Ray methods
Escherichia coli genetics
Escherichia coli metabolism
Molecular Chaperones genetics
Protein Subunits chemistry
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins isolation & purification
Static Electricity
Yersinia pestis genetics
Antigens, Bacterial chemistry
Bacterial Proteins chemistry
Molecular Chaperones chemistry
Yersinia pestis chemistry
Subjects
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