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

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Marcos Vicente de A S NavarroRichard C Garratt

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.

References

Mar 1, 1992·European Journal of Biochemistry·W Bode, R Huber
Jan 5, 1991·Journal of Molecular Biology·S OnestiD M Blow
Feb 10, 2000·Acta Crystallographica. Section D, Biological Crystallography·Z DauterK R Rajashankar
Jun 22, 2001·Acta Crystallographica. Section D, Biological Crystallography·R A NagemI Polikarpov
Apr 12, 1965·Biochimica Et Biophysica Acta·R F STEINER

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Citations

Aug 10, 2010·International Journal of Biological Macromolecules·Rosemeire A Silva-LuccaMaria Luiza V Oliva
Jul 17, 2007·Biochemical and Biophysical Research Communications·Daiane HansenMaria Luiza Vilela Oliva
Aug 8, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Dongwen ZhouAlexander Wlodawer

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