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. 2005 Sep 1;61(Pt 9):834-6.
doi: 10.1107/S1744309105024012. Epub 2005 Aug 31.

Cloning, recombinant production, crystallization and preliminary X-ray diffraction studies of a family 84 glycoside hydrolase from Clostridium perfringens

Elizabeth Ficko-Blean  1 , Alisdair B Boraston
Affiliations

Cloning, recombinant production, crystallization and preliminary X-ray diffraction studies of a family 84 glycoside hydrolase from Clostridium perfringens

Elizabeth Ficko-Blean et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

Abstract

Clostridium perfringens is a ubiquitous environmental organism that is capable of causing a variety of diseases in mammals, including gas gangrene and necrotic enteritis in humans. The activity of a secreted hyaluronidase, attributed to the NagH protein, contributes to the pathogenicity of this organism. The family 84 catalytic module of one of the three homologues of NagH found in C. perfringens (ATCC 13124) has been cloned. The 69 kDa catalytic module of NagJ, here called GH84C, was overproduced in Escherichia coli and purified by immobilized metal-affinity chromatography (IMAC). Crystals belonging to space group I222 or I2(1)2(1)2(1) with unit-cell parameters a = 130.39, b = 150.05, c = 155.43 A were obtained that diffracted to 2.1 A. Selenomethionyl crystals have also been produced, leading to the possibility of solving the phase problem by MAD using synchrotron radiation.

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Figures

Figure 1
Figure 1
Modular structure of GH84C. Numbers indicate the amino acid at the putative module boundaries. Amino acids 1–30 represent the N-terminal secretion signal. Amino acids 31–624 represent the GH84C catalytic module. Amino acids 625–767 represent a family 32 carbohydrate-binding module. Amino acids 768–909 bound an unknown module. Amino acids 910–1001 share a distant sequence identity with fibronectin type III repeats.
Figure 2
Figure 2
(a) Crystals of native GH84C catalytic module. (b) Crystals of selenomethionine-derivatized GH84C catalytic module.
See this image and copyright information in PMC

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