Crystallization and preliminary X-ray analysis of a novel Kunitz-type kallikrein inhibitor from Bauhinia bauhinioides

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 Å resolution using an in‐house X‐ray generator. The crystals of the...

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Bibliographic Details
Published in:	Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 61; no. 10; pp. 910 - 913
Main Authors:	Navarro, Marcos Vicente de A. S., Vierira, Débora F., Nagem, Ronaldo A. P., De Araújo, Ana Paula U., Oliva, Maria Luiza V., Garratt, Richard C.
Format:	Journal Article
Language:	English
Published:	5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01-10-2005 International Union of Crystallography
Subjects:	ANIONS Bauhinia Bauhinia - metabolism Bauhinia bauhinioides CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY CRYSTAL STRUCTURE CRYSTALLIZATION Crystallization Communications Crystallography, X-Ray CRYSTALS Diffusion disulfide bridges ESCHERICHIA COLI Escherichia coli - metabolism Iodides - chemistry Kallikreins - antagonists & inhibitors Kallikreins - chemistry Kunitz-type kallikrein inhibitors MATHEMATICAL SOLUTIONS Models, Statistical MONOMERS Peptides - chemistry Plant Proteins - chemistry Polyethylene Glycols - chemistry Recombinant Proteins - chemistry RESOLUTION Sodium Iodide - chemistry SOLUTIONS SOLVENTS SPACE GROUPS Temperature X-RAY DIFFRACTION X-Rays
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Summary:	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 Å resolution using an in‐house X‐ray generator. The crystals of the recombinant protein (rBbKI) belong to the orthorhombic space group P212121, with unit‐cell parameters a = 46.70, b = 64.14, c = 59.24 Å. 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 Å3 Da−1). Iodinated crystals were prepared and a derivative data set was also collected at 2.1 Å 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.
Bibliography:	ArticleID:AYF2ZA5115 ark:/67375/WNG-S6J38X74-Z istex:85E4ABCBC9BC46F61C7EECB9D39661816995D3E3 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1744-3091 1744-3091
DOI:	10.1107/S1744309105028496