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Crystallization and preliminary X-ray analysis of a novel Kunitz-type kallikrein inhibitor from Bauhinia bauhinioides.
- Source :
-
Acta crystallographica. Section F, Structural biology and crystallization communications [Acta Crystallogr Sect F Struct Biol Cryst Commun] 2005 Oct 01; Vol. 61 (Pt 10), pp. 910-3. Date of Electronic Publication: 2005 Sep 30. - Publication Year :
- 2005
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Abstract
- A Kunitz-type protease inhibitor (BbKI) found in Bauhinia bauhinioides seeds has been overexpressed in Escherichia coli and crystallized at 293 K using PEG 4000 as the precipitant. X-ray diffraction data have been collected to 1.87 A resolution using an in-house X-ray generator. The crystals of the recombinant protein (rBbKI) belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 46.70, b = 64.14, c = 59.24 A. Calculation of the Matthews coefficient suggests the presence of one monomer of rBbKI in the asymmetric unit, with a corresponding solvent content of 51% (VM = 2.5 A3 Da(-1)). Iodinated crystals were prepared and a derivative data set was also collected at 2.1 A resolution. Crystals soaked for a few seconds in a cryogenic solution containing 0.5 M NaI were found to be reasonably isomorphous to the native crystals. Furthermore, the presence of iodide anions could be confirmed in the NaI-derivatized crystal. Data sets from native and derivative crystals are being evaluated for use in crystal structure determination by means of the SIRAS (single isomorphous replacement with anomalous scattering) method.
- Subjects :
- Crystallography, X-Ray
Diffusion
Escherichia coli metabolism
Iodides chemistry
Models, Statistical
Polyethylene Glycols chemistry
Recombinant Proteins chemistry
Sodium Iodide chemistry
Temperature
X-Ray Diffraction
X-Rays
Bauhinia metabolism
Kallikreins antagonists & inhibitors
Kallikreins chemistry
Peptides chemistry
Plant Proteins chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1744-3091
- Volume :
- 61
- Issue :
- Pt 10
- Database :
- MEDLINE
- Journal :
- Acta crystallographica. Section F, Structural biology and crystallization communications
- Publication Type :
- Academic Journal
- Accession number :
- 16511193
- Full Text :
- https://doi.org/10.1107/S1744309105028496