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24-Oct-2019 11:21

Interestingly, there were no ions present corresponding to two, four, six, or eight Kdo residues.

These experiments were repeated with two additional independent batches of K1 CPS terminus.

To determine the anomeric configuration of the Kdo residues, the K1 and K5 termini were further analyzed by NMR spectroscopy.

In the H gradient correlation spectroscopy (g COSY) and gradient total correlation spectroscopy (g TOCSY) spectra from 1.81 to 1.92 ppm, there are two species in solution that contain signals consistent with Kdo methylene protons (Table 2 and Fig. Both have a difference of 0.5 ppm between the axial and equatorial H3 protons, consistent with β-linkages (29).

Although the correlation between the duplicated genes and the proposed terminal Kdo residue has been noted, it does not represent a unifying feature for all bacteria containing these CPS assembly systems because other examples (e.g., ) lack the duplication.

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Kps MTED functions are not confined to a given CPS repeat-unit structure, and one possible explanation of their broad substrate specificity is the presence of a conserved lipid terminus that may be recognized by the ABC transporter (3, 5, 14, 15).As a result, prior studies have investigated material released from CPS preparations treated with acid.Although acid hydrolysates yield information on individual components, they provide no insight into the linkage.To fully understand the assembly pathway, it is critical to know the precise nature of the lipid terminus and its linkage to the CPS glycan.To that end, we have determined the structure of the reducing terminus of CPSs from Structural characterization of a lipid terminus and its linkage region is not feasible with heterogeneous preparations containing high-molecular-mass CPS glycans.

Kps MTED functions are not confined to a given CPS repeat-unit structure, and one possible explanation of their broad substrate specificity is the presence of a conserved lipid terminus that may be recognized by the ABC transporter (3, 5, 14, 15).As a result, prior studies have investigated material released from CPS preparations treated with acid.Although acid hydrolysates yield information on individual components, they provide no insight into the linkage.To fully understand the assembly pathway, it is critical to know the precise nature of the lipid terminus and its linkage to the CPS glycan.To that end, we have determined the structure of the reducing terminus of CPSs from Structural characterization of a lipid terminus and its linkage region is not feasible with heterogeneous preparations containing high-molecular-mass CPS glycans.To identify the linkage of the Kdo residues, methylation analysis of the K1 hydrophobic moiety was performed using the Ciucanu and Kerek method with modifications to preserve the linkages of Neu Ac and Kdo (25-methyl-octitol were observed (ratio of 1:1.38), suggestive of an alternating polymer of 2,4- and 2,7-linked Kdo (Fig. A small amount of terminal Kdo was also detected, indicative of some hydrolysis in the sample.