Document Type
Article
Language
eng
Format of Original
13 p.
Publication Date
12-2003
Publisher
American Society for Biochemistry and Molecular Biology
Source Publication
Journal of Biological Chemistry
Source ISSN
0021-9258
Abstract
The O-antigen polysaccharide (OPS) of Rhizobium etli CE3 lipopolysaccharide (LPS) is linked to the core oligosaccharide via an N-acetylquinovosaminosyl (QuiNAc) residue. A mutant of CE3, CE166, produces LPS with reduced amounts of OPS, and a suppressed mutant, CE166α, produces LPS with nearly normal OPS levels. Both mutants are deficient in QuiNAc production. Characterization of OPS from CE166 and CE166α showed that QuiNAc was replaced by its 4-keto derivative, 2-acetamido-2,6-dideoxyhexosyl-4-ulose. The identity of this residue was determined by NMR and mass spectrometry, and by gas chromatography-mass spectrometry analysis of its 2-acetamido-4-deutero-2,6-dideoxyhexosyl derivatives produced by reduction of the 4-keto group using borodeuteride. Mass spectrometric and methylation analyses showed that the 2-acetamido-2,6-dideoxyhexosyl-4-ulosyl residue was 3-linked and attached to the core-region external Kdo III residue of the LPS, the same position as that of QuiNAc in the CE3 LPS. DNA sequencing revealed that the transposon insertion in strain CE166 was located in an open reading frame whose predicted translation product, LpsQ, falls within a large family of predicted open reading frames, which includes biochemically characterized members that are sugar epimerases and/or reductases. A hypothesis to be tested in future work is that lpsQ encodes UDP-2-acetamido-2,6-dideoxyhexosyl-4-ulose reductase, the second step in the synthesis of UDP-QuiNAc from UDP-GlcNAc.
Recommended Citation
Forsberg, Lennart Scott; Noel, K. Dale; Box, Jodie M.; and Carlson, Russell W., "Genetic Locus and Structural Characterization of the Biochemical Defect in the O-Antigenic Polysaccharide of the Symbiotically Deficient Rhizobium etli Mutant, CE166" (2003). Biological Sciences Faculty Research and Publications. 41.
https://epublications.marquette.edu/bio_fac/41
Comments
Accepted version. Journal of Biological Chemistry, Vol. 278, No. 51 (December 2003): 51347-51359. DOI. © 2003 American Society for Biochemistry and Molecular Biology. Used with permission.
This research was originally published in the Journal of Biological Chemistry. L. Scott Forsberg, K. Dale Noel, Jodie Box and Russell W. Carlson. "Genetic locus and structural characterization of the biochemical defect in the O-antigenic polysaccharide of the symbiotically deficient rhizobium etli mutant CE166." Journal of Biological Chemistry. 2003. Vol 278:51347-51359. © the American Society for Biochemistry and Molecular Biology.