tailieunhanh - Báo cáo khoa học: The HS:1 serostrain of Campylobacter jejuni has a complex teichoic acid-like capsular polysaccharide with nonstoichiometric fructofuranose branches and O-methyl phosphoramidate groups

Recently, the CPS biosynthetic loci for several strains ofCampylobacter jejuni were sequenced and revealed evidence for multiple mechanisms of structural variation. In this study, the CPS structure for the HS:1 serostrain of C. jejuni was determined using mass spectrometry and NMR at 600 MHz equipped with an ultra-sensitive cryogenically cooled probe. Ana-lysis of CPS purified using a mild enzymatic method revealed a teichoic acid-like [-4)-a-d-Galp-(1–2)-(R)-Gro-(1-P]n, repeating unit, where Gro is glycerol | ềFEBS Journal The HS 1 serostrain of Campylobacter jejuni has a complex teichoic acid-like capsular polysaccharide with nonstoichiometric fructofuranose branches and O-methyl phosphoramidate groups David J. McNally Harold C. Jarrell Jianjun Li Nam H. Khieu Evgeny Vinogradov Christine M. Szymanski and Jean-Robert Brisson Institute for BiologicalSciences NationalResearch Councilof Canada Ottawa Ontario Canada Keywords Campylobacter jejuni capsular polysaccharide CE-ESI-MS HR-MAS NMR phosphoramidate Correspondence . Brisson Institute for Biological Sciences National Research Council of Canada Ottawa Canada K1A0R6 Fax 1 613 9529092 Tel 1 613 9903244 E-mail Received 11 May 2005 revised 4 July 2005 accepted 11 July 2005 doi Recently the CPS biosynthetic loci for several strains of Campylobacter jejuni were sequenced and revealed evidence for multiple mechanisms of structural variation. In this study the CPS structure for the HS 1 serostrain of C. jejuni was determined using mass spectrometry and NMR at 600 MHz equipped with an ultra-sensitive cryogenically cooled probe. Analysis of CPS purified using a mild enzymatic method revealed a teichoic acid-like -4 -a-D-Galp- 1-2 - R -Gro- 1-P n repeating unit where Gro is glycerol. Two branches at C-2 and C-3 of galactose were identified as P-D-fructofuranoses substituted at C-3 with CH3OP O NH2 OR groups. Structural heterogeneity was due to nonstoichiometric glycosylation at C-3 of galactose and variable phosphoramidate groups. Identical structural features were found for cell-bound CPS on intact cells using proton homonuclear and 31P heteronuclear two-dimensional HR-MAS NMR at 500 MHz. In contrast spectroscopic data acquired for hot water phenol purified CPS was complicated by the hydrolysis and subsequent loss of labile groups during extraction. Collectively the results of this study established the importance of using sensitive isolation techniques and .

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