8V4H

X-ray structure of the NADP-dependent reductase from Campylobacter jejuni responsible for the synthesis of CDP-glucitol in the presence of CDP-glucitol

Summary for 8V4H

• Entry DOI: 10.2210/pdb8v4h/pdb
• Related: 5v4g
• Descriptor: Putative nucleotide sugar dehydratase, [[(2~{R},3~{S},4~{R},5~{R})-5-(4-azanyl-2-oxidanylidene-pyrimidin-1-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-oxidanyl-phosphoryl] [(2~{R},3~{R},4~{R},5~{S})-2,3,4,5,6-pentakis(oxidanyl)hexyl] hydrogen phosphate, CHLORIDE ION, ... (6 entities in total)
• Functional Keywords: capsular polysaccharide, short chain dehydrogenase, oxidoreductase
• Biological source: Campylobacter jejuni
• Total number of polymer chains: 2
• Total formula weight: 83512.10

Authors

Thoden, J.B.,Schumann, M.E.,Holden, H.M.,Raushel, F.M. (deposition date: 2023-11-29, release date: 2023-12-20, Last modification date: 2024-03-13)

Primary citation

Ghosh, M.K.,Narindoshvili, T.,Thoden, J.B.,Schumann, M.E.,Holden, H.M.,Raushel, F.M.
Biosynthesis of Cytidine Diphosphate-6-d-Glucitol for the Capsular Polysaccharides of Campylobacter jejuni.
Biochemistry, 63:699-710, 2024

PubMed Abstract: is a Gram-negative pathogenic bacterium commonly found in chickens and is the leading cause of human diarrheal disease worldwide. The various serotypes of produce structurally distinct capsular polysaccharides (CPSs) on the exterior surfaces of the cell wall. The capsular polysaccharide from serotype HS:5 is composed of a repeating sequence of d--d--heptose and d-glucitol-6-phosphate. We previously defined the pathway for the production of d--d--heptose in . Here, we elucidate the biosynthetic pathway for the assembly of cytidine diphosphate (CDP)-6-d-glucitol by the combined action of two previously uncharacterized enzymes. The first enzyme catalyzes the formation of CDP-6-d-fructose from cytidine triphosphate (CTP) and d-fructose-6-phosphate. The second enzyme reduces CDP-6-d-fructose with NADPH to generate CDP-6-d-glucitol. Using sequence similarity network (SSN) and genome neighborhood network (GNN) analyses, we predict that these pairs of proteins are responsible for the biosynthesis of CDP-6-d-glucitol and/or CDP-d-mannitol in the lipopolysaccharides (LPSs) and capsular polysaccharides in more than 200 other organisms. In addition, high resolution X-ray structures of the second enzyme are reported, which provide novel insight into the manner in which an open-chain nucleotide-linked sugar is harbored in an active site cleft.

PubMed: 38386885
DOI: 10.1021/acs.biochem.3c00706

Experimental method

X-RAY DIFFRACTION (2.2 Å)