In:
Acta Crystallographica Section F Structural Biology and Crystallization Communications, International Union of Crystallography (IUCr), Vol. 68, No. 11 ( 2012-11-01), p. 1409-1414
Abstract:
Obtaining crystals presented a bottleneck in the structural study of Anabaena cyanobacterial Ca 2+ -binding protein (CcbP). In this report, the promoting effect of Ellman's reagent [5,5′-dithiobis(2-nitrobenzoic acid); DTNB] on the crystallization of CcbP is described. CcbP contains one free cysteine. A quick and simple oxidation reaction with DTNB blocked the free cysteine in purified CcbP and generated a homogenous monomeric protein for crystallization. The crystal structure of DTNB-modified CcbP was determined by the single-wavelength anomalous diffraction method. Structure analysis indicated that DTNB modification facilitated crystallization of CcbP by inducing polar interactions in the crystal lattice. DTNB-mediated cysteine modification was demonstrated to have little effect on the overall structure and the Ca 2+ binding of CcbP. Thus, DTNB modification may provide a simple and general approach for protein modification to improve the success of crystallization screening.
Type of Medium:
Online Resource
ISSN:
1744-3091
DOI:
10.1107/S1744309112034938
Language:
Unknown
Publisher:
International Union of Crystallography (IUCr)
Publication Date:
2012
detail.hit.zdb_id:
2175956-X
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