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

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Elizabeth Ficko-Blean, Alisdair B Boraston

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.

References

Apr 22, 1999·Trends in Microbiology·L PetitM R Popoff
Oct 26, 1999·Acta Crystallographica. Section D, Biological Crystallography·J W Pflugrath
Jan 17, 2002·Proceedings of the National Academy of Sciences of the United States of America·Tohru ShimizuHideo Hayashi

❮ Previous
Next ❯

Citations

Jul 18, 2009·The Journal of Biological Chemistry·Melanie A HigginsAlisdair B Boraston
Feb 6, 2009·The Journal of Biological Chemistry·Elizabeth Ficko-BleanAlisdair B Boraston
Mar 17, 2006·The EMBO Journal·Francesco V RaoDaan M F van Aalten
Apr 9, 2015·Frontiers in Genetics·Louise E TailfordNathalie Juge
Dec 26, 2006·Nature Structural & Molecular Biology·Alicia Lammerts van BuerenAlisdair B Boraston
Mar 2, 2011·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Eun J Kim
Sep 23, 2006·The Journal of Biological Chemistry·Elizabeth Ficko-Blean, Alisdair B Boraston

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cardiac Glycosides

Cardiac glycosides are a diverse family of naturally derived compounds that bind to and inhibit na+/k+-atpase. Discover the latest research on cardiac glycosides heres.