Dr. Peng Xu <xup3
Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville, Bethesda, MD, 20892-0815, USA. Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic, Institute of Epidemiology, Faculty of Medicine, Comenius University, Spitalska 24, 81372, Bratislava, Slovak Republic, Infectious Diseases Division, International Centre for, Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, 02114, USA, Harvard Medical School, Boston, MA, USA, Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA, USA
The lipopolysaccharide (LPS) of Vibrio cholerae O139, strain CIRS245, was isolated conventionally, and the lipid A was removed by mild acid hydrolysis (0.1 m NaOAc buffer containing 1 % SDS, pH 4.2, 95 �C, 8 h). The crude product was a complex mixture consisting mainly of constituent fragments of the O-specific polysaccharide-core (OSPc). The OSPc was only a minor component in the mixture. Two-stage purification of the crude OSPc by HPLC gave pure OSPc fragment of the LPS, as shown by NMR spectroscopy, analytical HPLC and ESI-MS. This material is the purest OSPc fragment of the LPS from Vibrio cholerae O139 reported to date. The purified OSPc was readily converted to the corresponding methyl squarate derivative and the latter was conjugated to BSA. The conjugate, when examined by ELISA, showed immunoreactivity with sera from patients in Bangladesh recovering from cholera caused by V. cholerae O139, but not O1.
13C NMR, 1H NMR, NMR-2D, SDS-PAGE, ELISA, ESI-MS, mild acid hydrolysis, MALDI-TOF MS, HPLC, SEC, conjugation, fluorescence binding assay, immunoreactivity assays
the structure of the delipidated LPS of V. cholerae O139.
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6 C7 C8
5,3,2,3,3,4,3,2 aXColp 99.26 63.41 32.62 68.60 66.02 15.30
5,3,2,3,3,4,3,6 P
5,3,2,3,3,4,3 bDGalp 100.94 75.89 72.32 76.28 67.25 68.55
5,3,2,3,3,4,2 Ac 174.02 22.39
5,3,2,3,3,4,4 aXColp 97.62 63.20 32.47 68.28 66.57 15.43
5,3,2,3,3,4 bDGlcpN 102.92 55.59 75.38 72.55 75.33 59.95
5,3,2,3,3 aDGalpA 101.63 68.17 68.84 78.69 70.87 173.29
5,3,2,3,2 Ac 174.07 22.07
5,3,2,3 bDQuipN 101.63 54.47 81.11 75.63 71.58 16.57
5,3,2,2 Ac 172.82 20.27
5,3,2,7 aDGlcpN 96.24 54.11 69.93 69.13 72.72 60.05
5,3,2 aXLDmanHepp 98.40 70.67 79.19 65.12 72.25 67.57 70.61
5,3,6 aXLDmanHepp 98.68 70.91 70.76 65.57 72.11 68.99 62.47
5,3 aXLDmanHepp 98.76 79.41 69.73 66.32 71.52 73.10 62.41
5,4 bDGlcp 102.35 73.80 75.94 70.31 75.98 60.64
5,6 aDGlcp 101.43 71.84 72.78 68.70 71.66 59.71
5 aXLDmanHepp 101.81 70.06 73.40 73.95 71.55 79.09 61.54
xX4,8anhKdoa 169.06 203.30 41.97 74.04 79.75 73.99 69.38 69.85
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6 H7 H8
5,3,2,3,3,4,3,2 aXColp 4.92 3.87 1.78 3.69 4.19 1.10
5,3,2,3,3,4,3,6 P
5,3,2,3,3,4,3 bDGalp 4.62 3.52 3.81 4.47 3.52 4.23-4.31
5,3,2,3,3,4,2 Ac - 1.96
5,3,2,3,3,4,4 aXColp 4.77 3.90 1.76-1.96 4.11 4.67 1.10
5,3,2,3,3,4 bDGlcpN 4.39 3.74 3.92 3.57 3.32 3.72-3.81
5,3,2,3,3 aDGalpA 5.30 3.57 3.82 4.16 4.08 -
5,3,2,3,2 Ac - 1.84
5,3,2,3 bDQuipN 4.55 3.63 3.51 3.34 3.43 1.23
5,3,2,2 Ac - 2.05
5,3,2,7 aDGlcpN 4.97 3.37 3.81 3.51 3.61 3.71-3.79
5,3,2 aXLDmanHepp 4.99 4.99 3.95 3.87 3.61 4.17 3.51-3.86
5,3,6 aXLDmanHepp 5.16 3.74 3.67 3.80 3.56 3.93 3.64-3.68
5,3 aXLDmanHepp 5.42 4.12 3.88 3.73 3.63 4.05 3.69-3.90
5,4 bDGlcp 4.52 3.13 3.37 3.02 3.28 3.64-3.82
5,6 aDGlcp 5.08 3.45 3.68 3.40 3.80 3.72-3.76
5 aXLDmanHepp 5.05 4.05 3.78 4.14 3.71 4.00 3.65-3.78
xX4,8anhKdoa - - 3.04 3.58 3.53 3.63 3.84 3.52-3.74
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6 C7/H7 C8/H8
5,3,2,3,3,4,3,2 aXColp 99.26/4.92 63.41/3.87 32.62/1.78 68.60/3.69 66.02/4.19 15.30/1.10
5,3,2,3,3,4,3,6 P
5,3,2,3,3,4,3 bDGalp 100.94/4.62 75.89/3.52 72.32/3.81 76.28/4.47 67.25/3.52 68.55/4.23-4.31
5,3,2,3,3,4,2 Ac 22.39/1.96
5,3,2,3,3,4,4 aXColp 97.62/4.77 63.20/3.90 32.47/1.76-1.96 68.28/4.11 66.57/4.67 15.43/1.10
5,3,2,3,3,4 bDGlcpN 102.92/4.39 55.59/3.74 75.38/3.92 72.55/3.57 75.33/3.32 59.95/3.72-3.81
5,3,2,3,3 aDGalpA 101.63/5.30 68.17/3.57 68.84/3.82 78.69/4.16 70.87/4.08
5,3,2,3,2 Ac 22.07/1.84
5,3,2,3 bDQuipN 101.63/4.55 54.47/3.63 81.11/3.51 75.63/3.34 71.58/3.43 16.57/1.23
5,3,2,2 Ac 20.27/2.05
5,3,2,7 aDGlcpN 96.24/4.97 54.11/3.37 69.93/3.81 69.13/3.51 72.72/3.61 60.05/3.71-3.79
5,3,2 aXLDmanHepp 98.40/4.99 70.67/4.99 79.19/3.95 65.12/3.87 72.25/3.61 67.57/4.17 70.61/3.51-3.86
5,3,6 aXLDmanHepp 98.68/5.16 70.91/3.74 70.76/3.67 65.57/3.80 72.11/3.56 68.99/3.93 62.47/3.64-3.68
5,3 aXLDmanHepp 98.76/5.42 79.41/4.12 69.73/3.88 66.32/3.73 71.52/3.63 73.10/4.05 62.41/3.69-3.90
5,4 bDGlcp 102.35/4.52 73.80/3.13 75.94/3.37 70.31/3.02 75.98/3.28 60.64/3.64-3.82
5,6 aDGlcp 101.43/5.08 71.84/3.45 72.78/3.68 68.70/3.40 71.66/3.80 59.71/3.72-3.76
5 aXLDmanHepp 101.81/5.05 70.06/4.05 73.40/3.78 73.95/4.14 71.55/3.71 79.09/4.00 61.54/3.65-3.78
xX4,8anhKdoa 41.97/3.04 74.04/3.58 79.75/3.53 73.99/3.63 69.38/3.84 69.85/3.52-3.74
report error
Found 1 record.
Displayed record 1
, ICD11: XN7N1 
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