Crystal structure of Bacillus anthracis transpeptidase enzyme CapD

Abstract

Bacillus anthracis elaborates a poly-γ-D-glutamic acid capsule that protects bacilli from phagocytic killing during infection. The enzyme CapD generates amide bonds with peptidoglycan cross-bridges to anchor capsular material within the cell wall envelope of B. anthracis. The capsular biosynthetic pathway is essential for virulence during anthrax infections and can be targeted for anti-infective inhibition with small molecules. Here, we present the crystal structures of the γ-glutamyltranspeptidase CapD with and without α-L-Glu-L-Glu dipeptide, a non-hydrolyzable analog of poly-γ-D-glutamic acid, in the active site. Purified CapD displays transpeptidation activity in vitro, and its structure reveals an active site broadly accessible for poly-γ-glutamate binding and processing. Using structural and biochemical information, we derive a mechanistic model for CapD catalysis whereby Pro427, Gly428, and Gly429 activate the catalytic residue of the enzyme, Thr352, and stabilize an oxyanion hole via main chain amide hydrogen bonds.