Expression and crystallographic studies of d‐glycero‐β‐d‐manno‐heptose‐1‐phosphate adenylyltransferase from Burkholderia pseudomallei. Issue 2 (1st February 2017)

• Record Type: Journal Article
• Authors: Park, Jimin; Kim, Hyojin; Kim, Suwon; Lee, Daeun; Shin, Dong Hae
• Is Part Of: Acta crystallographica. Volume 73:Issue 2(2017:Feb.)
• Page Start: 90
• Page End: 94
• Publication Date: 2017-02-01
• Subjects: Burkholderia pseudomallei -- heptose biosynthesis pathway -- melioidosis -- d‐glycero‐β‐d‐manno‐heptose‐1‐phosphate adenylyltransferase -- HldC; Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X16020537
• Languages: English
• ISSNs: 2053-230X

Abstract

Abstract : d ‐ glycero ‐β‐d ‐ manno ‐Heptose‐1‐phosphate adenylyltransferase (HldC) from B. pseudomallei has been cloned, expressed, purified and crystallized. Synchrotron X‐ray data from a selenomethionine‐substituted HldC crystal were collected to 2.8 Å resolution. Abstract : 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 P 1, 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.