Table 2.
Negative-ion CE-ES-MS data and proposed compositions of O-deacylated LPS (LPS-OH) and core OS from the Heddleston serovar 3 and 4 type strains (P1059 and P1662), and proposed compositions of LPS-OH from the P1059 TargeTron mutants
| Study and serovar (description) | Observed ions (m/z) |
Molecular mass (Da)a |
Relative intensity | Proposed composition | ||||
|---|---|---|---|---|---|---|---|---|
| (M − H)− | (M − 4H)4− | (M − 3H)3− | (M − 2H)2− | Observed | Calculated | |||
| Serovar study | ||||||||
| Serovar 3 (P1059) | 1,106.5 | 3,322 | 3,320.0 | 0.2 | HexNAc, 6Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | |||
| LPS-OH | 1,147.3 | 1,721.2 | 3,444 | 3,443.0 | 1.0 | HexNAc, 6Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||
| 889.8 | 1,188.2 | 3,568 | 3,566.0 | 0.8 | HexNAc, 6Hex, 4Hep, Kdo-P, 3PEtn, lipid A-OH | |||
| Serovar 4 (P1662) | ||||||||
| LPS-OH | 707.4 | 943.5 | 2,833.6 | 2,831.6 | 0.2 | 5Hex, 4Hep, Kdo-P, lipid A-OH | ||
| 738.0 | 984.3 | 1,477.2 | 2,956.0 | 2,954.7 | 0.5 | 5Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | ||
| 768.6 | 1,025.4 | 1,538.7 | 3,079.0 | 3,077.7 | 1.0 | 5Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||
| 799.5 | 1,066.2 | 3,202.2 | 3,200.7 | 0.5 | 5Hex, 4Hep, Kdo-P, 3PEtn, lipid A-OH | |||
| Core OS | 899.1 | 1,800.2 | 1,799.5 | 0.2 | 5Hex, 4Hep, aKdob | |||
| 960.3 | 1,922.6 | 1,922.6 | 0.8 | 5Hex, 4Hep, PEtn, aKdob | ||||
| 1,022.1 | 2,046.2 | 2,045.7 | 1.0 | 5Hex, 4Hep, 2PEtn, aKdob | ||||
| GctC study | ||||||||
| AL539 | ||||||||
| LPS-OH (gatA polar mutant) | 709.7 | 1,064.9 | 2,131.9 | 2,130.9 | 0.2 | Hex, 3Hep, 2Kdo, lipid A-OH | ||
| 717.0 | 1,075.9 | 2,153.9 | 2,152.9 | 1.0 | 2Hex, 3Hep, Kdo-P, lipid A-OH | |||
| LPS-OH | 758.0 | 1,137.4 | 2,276.9 | 2,276.0 | 0.6 | 2Hex, 3Hep, Kdo-P, PEtn, lipid A-OH | ||
| 773.5 | 1,160.9 | 2,323.7 | 2,323.1 | 0.1 | Hex, 4Hep, 2Kdo, lipid A-OH | |||
| 780.5 | 1,171.5 | 2,344.8 | 2,345.1 | 0.2 | 2Hex, 4Hep, Kdo-P, lipid A-OH | |||
| 822.1 | 1,233.2 | 2,469.2 | 2,468.2 | 0.1 | 2Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | |||
| AL806 LPS-OH (AL539 + gctC, hptE) | 691.8 | 923.2 | 2,772.1 | 2,770.5 | 0.5 | 3Hex, 4Hep, 2Kdo, PEtn, lipid A-OH | ||
| 930.4 | 2,794.2 | 2,792.4 | 0.3 | 4Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | ||||
| 728.5 | 971.4 | 2,917.6 | 2,915.5 | 1.0 | 4Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | |||
| GatF study | ||||||||
| AL2116 LPS-OH (gatF mutant) | 697.7 | 930.7 | 1,395.5 | 2,794.0 | 2,792.4 | 0.3 | 4Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | |
| 728.3 | 971.2 | 1,457.2 | 2,916.6 | 2,915.5 | 1.0 | 4Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||
| AL2297 LPS-OH (gatF mutant + gatF) | 1,174.0 | 3,525.0 | 3,523.2 | 0.4 | 2HexNAc, 6Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | |||
| 1,215.0 | 3,648.0 | 3,646.3 | 1.0 | 2HexNAc, 6Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||||
| GatG study | ||||||||
| AL2155 LPS-OH (gatG mutant) | 768.9 | 1,025.3 | 3,079.2 | 3,077.7 | 1.0 | 5Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||
| 799.6 | 1,066.2 | 3,202.0 | 3,200.8 | 1.0 | 5Hex, 4Hep, Kdo-P, 3PEtn, lipid A-OH | |||
| AL2192 LPS-OH (gatG mutant + gatG) | 1,106.5 | 3,322.5 | 3,320.0 | 1.0 | HexNAc, 6Hex, 4Hep, Kdo-P, lipid A-OH | |||
| 1,147.5 | 3,445.5 | 3,443.1 | 0.9 | HexNAc, 6Hex, 4Hep, Kdo-P, lipid A-OH | ||||
| 1,174.0 | 3,525.0 | 3,523.2 | 0.3 | 2HexNAc, 6Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | ||||
| 1,215.0 | 3,648.0 | 3,646.3 | 0.4 | 2HexNAc, 6Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||||
| NatB study | ||||||||
| AL2117 LPS-OH (natB mutant) | 768.7 | 1,025.4 | 3,079.0 | 3,077.7 | 0.8 | 5Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||
| 809.4 | 1,079.5 | 3,241.5 | 3,239.9 | 1.0 | 6Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | |||
| AL2299 LPS-OH (natB mutant + natB) | 1,174.0 | 3,525.0 | 3,523.2 | 0.4 | 2HexNAc, 6Hex, 4Hep, Kdo-P, PEtn, lipid A-OH | |||
| 1,215.0 | 3,648.0 | 3,646.3 | 1.0 | 2HexNAc, 6Hex, 4Hep, Kdo-P, 2PEtn, lipid A-OH | ||||
a
Average mass units were used for calculation of molecular mass based on the proposed composition, as follows: lipid A-OH, 952.00; Hex, 162.15; Hep, 192.17; Kdo, 220.18; anhydro Kdo (aKdo), 202.18; PEtn, 123.05; P, 79.95.
b
aKdo, hydrolysis products of Kdo formed during core hydrolysis.