TY - JOUR
T1 - Structure and serological characterization of the O-antigen of Proteus mirabilis O18 with a phosphocholine-containing oligosaccharide phosphate repeating unit
AU - Fudala, Rafal
AU - Kondakova, Anna N.
AU - Bednarska, Katarzyna
AU - Senchenkova, Sof'ya N.
AU - Shashkov, Alexander S.
AU - Knirel, Yuriy A.
AU - Zähringer, Ulrich
AU - Kaca, Wieslaw
N1 - Funding Information:
Authors thank Dr B. Lindner for providing access to the ion cyclotron resonance Fourier transform mass spectrometer and O.V. Bystrova for technical assistance. This work was supported by the Russian Foundation for Basic Research (grant 02-04-48767), INTAS (grant YSF 2001-2/1), and the Sciences Research Committee (KBN, Poland; grant.6PO4C 056 20).
PY - 2003/9/1
Y1 - 2003/9/1
N2 - A phosphorylated, choline-containing polysaccharide was obtained by O-deacylation of the lipopolysaccharide (LPS) of Proteus mirabilis O18 by treatment with aqueous 12% ammonia, whereas hydrolysis with dilute acetic acid resulted in depolymerisation of the polysaccharide chain by the glycosyl phosphate linkage. Treatment of the O-deacylated LPS with aqueous 48% hydrofluoric acid cleaved the glycosyl phosphate group but, unexpectedly, did not affect the choline phosphate group. The polysaccharide and the derived oligosaccharides were studied by NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C HMQC and HMQC-TOSCY experiments, along with chemical methods, and the following structure of the pentasaccharide phosphate repeating unit was established:Where ChoP=Phosphocoline Immunochemical studies of the LPS, O-deacylated LPS and partially dephosphorylated pentasaccharide using rabbit polyclonal anti-P. mirabilis O18 serum showed the importance of the glycosyl phosphate group in manifesting the serological specificity of the O18-antigen.
AB - A phosphorylated, choline-containing polysaccharide was obtained by O-deacylation of the lipopolysaccharide (LPS) of Proteus mirabilis O18 by treatment with aqueous 12% ammonia, whereas hydrolysis with dilute acetic acid resulted in depolymerisation of the polysaccharide chain by the glycosyl phosphate linkage. Treatment of the O-deacylated LPS with aqueous 48% hydrofluoric acid cleaved the glycosyl phosphate group but, unexpectedly, did not affect the choline phosphate group. The polysaccharide and the derived oligosaccharides were studied by NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C HMQC and HMQC-TOSCY experiments, along with chemical methods, and the following structure of the pentasaccharide phosphate repeating unit was established:Where ChoP=Phosphocoline Immunochemical studies of the LPS, O-deacylated LPS and partially dephosphorylated pentasaccharide using rabbit polyclonal anti-P. mirabilis O18 serum showed the importance of the glycosyl phosphate group in manifesting the serological specificity of the O18-antigen.
KW - Glycosyl phosphate
KW - Lipopolysaccharide
KW - O-Polysaccharide
KW - Phosphocholine
KW - Proteus mirabilis
UR - http://www.scopus.com/inward/record.url?scp=0142090511&partnerID=8YFLogxK
U2 - 10.1016/S0008-6215(03)00274-X
DO - 10.1016/S0008-6215(03)00274-X
M3 - Article
C2 - 12932366
AN - SCOPUS:0142090511
SN - 0008-6215
VL - 338
SP - 1835
EP - 1842
JO - Carbohydrate Research
JF - Carbohydrate Research
IS - 18
ER -