Protein expression, characterization, crystallization and preliminary X-ray crystallographic analysis of a Fic protein from Clostridium difficile.

ABSTRACT: Fic domains in proteins are found in abundance in nature from the simplest prokaryotes to animals. Interestingly, Fic domains found in two virulence factors of Gram-negative bacteria have recently been demonstrated to catalyse the transfer of the AMP moiety from ATP to small host GTPases. This post-translational modification has attracted considerable interest and a role for adenylylation in pathology and physiology is emerging. This work was aimed at the structural characterization of a newly identified Fic protein of the Gram-positive bacterium Clostridium difficile. A constitutively active inhibitory helix mutant of C. difficile Fic was overexpressed in Escherichia coli, purified and crystallized by the vapour-diffusion technique. Preliminary X-ray crystallographic analysis shows that the crystals diffract to at least 1.68?Å resolution at a synchrotron X-ray source. The crystals belonged to the orthorhombic space group P2?2?2?, with unit-cell parameters a=45.6, b=80.8, c=144.7?Å, ?=?=?=90°. Two molecules per asymmetric unit corresponds to a Matthews coefficient of 2.37?Å3?Da(-1) and a solvent content of 48%.

SUBMITTER: Welner D 

PROVIDER: S-EPMC4051547 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature