Expression and crystallographic studies of D-glycero-β-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.
College of Pharmacy, Ewha W. University, 52 Ewhayeodae-gil, Seoul 03760, Republic of Korea.
Jimin
J
Park
College of Pharmacy, Ewha W. University, 52 Ewhayeodae-gil, Seoul 03760, Republic of Korea.
Hyojin
H
Kim
College of Pharmacy, Ewha W. University, 52 Ewhayeodae-gil, Seoul 03760, Republic of Korea.
Suwon
S
Kim
College of Pharmacy, Ewha W. University, 52 Ewhayeodae-gil, Seoul 03760, Republic of Korea.
Daeun
D
Lee
College of Pharmacy, Ewha W. University, 52 Ewhayeodae-gil, Seoul 03760, Republic of Korea.
Dong Hae
DH
Shin
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016957:Burkholderia pseudomallei / Q000737:chemistry* / Q000201:enzymology; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006539:Heptoses / Q000737:chemistry* / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D006539:Heptoses; D011994:Recombinant Proteins; C026312:glycero-manno-heptose; D017853:Phosphotransferases (Alcohol Group Acceptor)
Burkholderia pseudomallei; HldC; d-glycero-β-d-manno-heptose-1-phosphate adenylyltransferase; heptose biosynthesis pathway; melioidosis