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Title: Purification, crystallization and preliminary X-ray diffraction analysis of the carbohydrate-binding region of the Streptococcus gordonii adhesin GspB

Purification, crystallization and preliminary X-ray diffraction analysis of the carbohydrate-binding region of the Streptococcus gordonii adhesin GspB The carbohydrate-binding region of the bacterial adhesin GspB from Streptococcus gordonii strain M99 (GspB{sub BR}) was expressed in Escherichia coli and purified using affinity and size-exclusion chromatography. Separate sparse-matrix screening of GspB{sub BR} buffered in either 20 mM Tris pH 7.4 or 20 mM HEPES pH 7.5 resulted in different crystallographic behavior such that different precipitants, salts and additives supported crystallization of GspB{sub BR} in each buffer. While both sets of conditions supported crystal growth in space group P2{sub 1}2{sub 1}2{sub 1}, the crystals had distinct unit-cell parameters of a = 33.3, b = 86.7, c = 117.9 {angstrom} for crystal form 1 and a = 34.6, b = 98.3, c = 99.0 {angstrom} for crystal form 2. Additive screening improved the crystals grown in both conditions such that diffraction extended to beyond 2 {angstrom} resolution. A complete data set has been collected to 1.3 {angstrom} resolution with an overall R{sub merge} value of 0.04 and an R{sub merge} value of 0.33 in the highest resolution shell.
Authors: ; ; ; ; ; ; ;
Publication Date:
OSTI Identifier:OSTI ID: 1037901
Resource Type:Journal Article
Resource Relation:Journal Name: Acta Crystallogr. F; Journal Volume: 66; Journal Issue: (11) ; 11, 2010
Research Org:Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
Sponsoring Org:OTHERAHANIH
Country of Publication:United States
Language:ENGLISH
Subject: 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; ADDITIVES; AFFINITY; CHROMATOGRAPHY; CRYSTAL GROWTH; CRYSTALLIZATION; DIFFRACTION; ESCHERICHIA COLI; GLYCOPROTEINS; LECTINS; PURIFICATION; RESOLUTION; SIALIC ACID; SPACE GROUPS; STRAINS; STREPTOCOCCUS; X-RAY DIFFRACTION