Structural characterisation 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 an AMP moiety from ATP to small host GTPases (1,2). This post-translational modification has received considerable interest and a role for adenylylation in pathology and physiology is emerging. We have structurally characterised a newly identified Fic protein of the pathogenic gram-positive bacterium Clostridium difficile. A constitutively active inhibitory helix mutant of C. difficile Fic was purified, crystallised and data collected to 1.7 Å resolution. The structure confirms C. difficult Fic protein as an ATP binding protein and reveal a binding site similar to other confirmed virulent Fic proteins. Surprisingly, this gram-positive Fic protein does not seem to target GTPases in humans and currently target identification is being chased. The current status of the project will be presented.