Expression and crystallographic studies of d-glycero-β-d-manno-heptose-1-phosphate adenylyltransferase from Burkholderia pseudomallei
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- 作者：Jimin Park ; Hyojin Kim ; Suwon Kim ; Daeun Lee and Dong Hae Shin
- 关键词：Burkholderia pseudomallei ; heptose biosynthesis pathway ; melioidosis ; d-glycero-&beta ; -d-manno-heptose-1-phosphate adenylyltransferase ; HldC
- 刊名：Acta Crystallographica Section F
- 出版年：2017
- 出版时间：February 2017
- 年：2017
- 卷：73
- 期：2
- 页码：90-94
- 全文大小：666K
- ISSN：1744-3091
文摘
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.