Dipartimento di Scienze Chimiche, Universita di Napoli Federico II, Complesso Universitario Monte S. Angelo, Via Cintia 4, 80126 Napoli (Italy)
This is the first report of the chemical and biological properties of the lipooligosaccharide (LOS) endotoxin isolated from Burkholderia dolosa IST4208, an isolate recovered from a cystic fibrosis (CF) patient in a Portuguese CF center. B. dolosa is a member of the Burkholderia cepacia complex, a group of closely related species that are highly problematic and opportunistic pathogens in CF. B. dolosa infection leads to accelerated loss of lung function and decreased survival. The structural determination of its endotoxin was achieved using a combination of chemistry and spectroscopy, and has revealed a novel endotoxin structure. The purified LOS was tested for its immunostimulatory activity on human HEK 293 cells expressing TLR-4, MD-2, and CD-14. In these assays, the LOS showed strong proinflammatory activity.
13C NMR, 1H NMR, NMR-2D, GC-MS, SDS-PAGE, sugar analysis, ELISA, mild acid hydrolysis, MALDI-TOF MS, NMR-1D, methanolysis, statistical analysis
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6 C7 C8
4 aXKop ? ? 70.8 71.6 ? 71.8 71.9 62.8
5,3,2 bDGlcp 101.3 72.2 75.8 71.6 76.4 60.8
5,3,3,2,3 aXLDmanHepp 102.3 69.8 69.3 71.1 71.1 67.9 62.9
5,3,3,2,7,2 Ac
5,3,3,2,7 bDQuipN 100.6 54.1 74.7 75.6 75.4 16.2
5,3,3,2 aXLDmanHepp 97.4 70.7 76.5 71.5 72.2 70.4 70.1
5,3,3,6,3 bDGalp 103.3 71.1 72.0 68.5 75.2 60.5
5,3,3,6 aDGlcp 97.6 71.1 82.5 72.9 68.0 60.3
5,3,3 aDGalp 97.3 74.4 71.3 70.3 71.2 64.6
5,3,7 aXLDmanHepp 100.6 69.9 70.4 71.1 71.1 68.7 62.9
5,3 aXLDmanHepp 99.1 77.2 79.9 68.9 70.7 71.9 70.1
5,4 bDGlcp 102.0 73.4 75.7 68.9 76.3 60.8
5 aXLDmanHepp 99.1 65.7 71.6 71.6 72.1 68.7 63.0
aXKdo? 174.1 ? 33.4 71.9 69.1 71.6 69.3 62.7
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6 H7 H8
4 aXKop - - 3.70 3.93 ? 3.56 3.99 3.59-3.86
5,3,2 bDGlcp 4.30 3.26 3.58 3.40 3.35 3.64
5,3,3,2,3 aXLDmanHepp 4.95 4.00 4.11 3.64 3.53 4.07 3.63
5,3,3,2,7,2 Ac
5,3,3,2,7 bDQuipN 4.50 3.69 3.73 3.24 3.40 1.21
5,3,3,2 aXLDmanHepp 4.97 3.88 4.03 3.70 3.53 3.95 3.55-3.63
5,3,3,6,3 bDGalp 4.56 3.49 3.60 3.82 3.64 3.67
5,3,3,6 aDGlcp 4.90 3.67 3.85 3.44 3.44 3.76
5,3,3 aDGalp 5.35 3.56 3.73 3.99 3.64 3.57-4.00
5,3,7 aXLDmanHepp 4.82 3.88 3.76 3.64 3.53 3.91 3.62
5,3 aXLDmanHepp 5.20 4.08 3.91 3.55 3.70 3.64 3.55-3.63
5,4 bDGlcp 4.46 3.22 3.58 3.56 3.35 3.64
5 aXLDmanHepp 5.19 3.95 3.93 3.97 3.81 3.91 3.49-3.92
aXKdo? - - 1.85-2.28 4.02 4.16 3.71 3.78 3.58-3.87
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6 C7/H7 C8/H8
4 aXKop 70.8/3.70 71.6/3.93 ?/? 71.8/3.56 71.9/3.99 62.8/3.59-3.86
5,3,2 bDGlcp 101.3/4.30 72.2/3.26 75.8/3.58 71.6/3.40 76.4/3.35 60.8/3.64
5,3,3,2,3 aXLDmanHepp 102.3/4.95 69.8/4.00 69.3/4.11 71.1/3.64 71.1/3.53 67.9/4.07 62.9/3.63
5,3,3,2,7,2 Ac
5,3,3,2,7 bDQuipN 100.6/4.50 54.1/3.69 74.7/3.73 75.6/3.24 75.4/3.40 16.2/1.21
5,3,3,2 aXLDmanHepp 97.4/4.97 70.7/3.88 76.5/4.03 71.5/3.70 72.2/3.53 70.4/3.95 70.1/3.55-3.63
5,3,3,6,3 bDGalp 103.3/4.56 71.1/3.49 72.0/3.60 68.5/3.82 75.2/3.64 60.5/3.67
5,3,3,6 aDGlcp 97.6/4.90 71.1/3.67 82.5/3.85 72.9/3.44 68.0/3.44 60.3/3.76
5,3,3 aDGalp 97.3/5.35 74.4/3.56 71.3/3.73 70.3/3.99 71.2/3.64 64.6/3.57-4.00
5,3,7 aXLDmanHepp 100.6/4.82 69.9/3.88 70.4/3.76 71.1/3.64 71.1/3.53 68.7/3.91 62.9/3.62
5,3 aXLDmanHepp 99.1/5.20 77.2/4.08 79.9/3.91 68.9/3.55 70.7/3.70 71.9/3.64 70.1/3.55-3.63
5,4 bDGlcp 102.0/4.46 73.4/3.22 75.7/3.58 68.9/3.56 76.3/3.35 60.8/3.64
5 aXLDmanHepp 99.1/5.19 65.7/3.95 71.6/3.93 71.6/3.97 72.1/3.81 68.7/3.91 63.0/3.49-3.92
aXKdo? 33.4/1.85-2.28 71.9/4.02 69.1/4.16 71.6/3.71 69.3/3.78 62.7/3.58-3.87
report error
Found 1 record.
Displayed record 1
]; pneumonia [ICD11: CA40 
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