Expression and crystallographic studies of D-glycero-b-D-manno-heptose-1-phosphate adenylyltransferase from Burkholderia pseudomallei

The Gram negative bacterium Burkholderia pseudomallei is the causative agent of melioidosis. d glycero β d manno Heptose 1 phosphate adenylyltransferase (HldC) is the fourth enzyme of the ADP l glycero β d manno heptose biosynthesis pathway, which produces an essential carbohydrate comprising the inner core of lipopolysaccharide. Therefore, HldC is a potential target of antibiotics against melioidosis. In this study, HldC from B. pseudomallei has been cloned, expressed, purified and crystallized. Synchrotron X ray data from a selenomethionine substituted HldC crystal were also collected to 2.8 Å resolution. The crystal belonged to the primitive triclinic space group P1, with unit cell parameters a = 74.0, b = 74.0, c = 74.9 Å, α = 108.4, β = 108.4, γ = 108.0°. Eight protomers are present in the unit cell and three out of five selenomethionines were found in each protomer using the PHENIX software suite. A full structural determination is in progress to elucidate the structure-function relationship of the protein.