TABLE 1.
B. subtilis muropeptidea | B. anthracis muropeptide | Structureb | Dormant sporec | Germinated spore pelletc | Germinated spore exudate after mutanolysin digestionc | Germinated spore exudate before mutanolysin digestionc | Cochromatography | m/z expectedd | m/z observedd | Amino acid/amino sugar analysise
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Muramic acid derived from NAM or muramic-δ-lactam | Glucosamine | Muramitol | Glucosaminitol | Ala | Glu | Dpm | ||||||||||
Muropeptides produced from dormant spore PG | ||||||||||||||||
1 | A | DS-TriP | + | * | + | + | + | 1 | 1 | 1 | ||||||
B | DS-Ac-TriP | + | + | 827.4 | 827.3 | + | + | 1 | 1 | 1 | ||||||
C | DS-TriP+Am | + | + | 868.7 | 869.7 | + | + | 1 | 1 | 1 | ||||||
D | DS-Ac-TriP+Am | + | + | 826.4 | 826.3 | + | + | 1 | 1 | 1 | ||||||
2 | E | DS-Ala | + | * | + | + | + | + | + | 1 | ||||||
3 | F | DS-TP | + | * | + | + | + | + | + | 2 | 1 | 1 | ||||
G | DS-Ac-TP | + | + | 898.4 | 898.3 | + | + | 2 | 1 | 1 | ||||||
5 | H | TS-TP open lactam | + | * | + | + | + | + | 2 | 1 | 1 | |||||
6 | I | TSred-TP | + | * | + | + | + | 2 | 1 | 1 | ||||||
7 | J | TSred-Ala | + | * | + | + | + | 1 | ||||||||
K | DS-Ac-TP x TriP+Am-DS-Ac | + | + | 1707.8 | 1707.7 | + | + | 3 | 2 | 2 | ||||||
L | TS-TP-Ac | + | * | + | 1316.5 | 1317.8 | + | + | + | 2 | 1 | 1 | ||||
M | DS-Ac-TP+Am x TriP+Am-DS-Ac | + | * | + | + | |||||||||||
10 | N | TS-TP | + | + | + | + | 1358.6 | 1358.5 | + | + | + | 2 | 1 | 1 | ||
11 | O | TS-TP x TP | + | * | + | + | + | + | 2 | 1 | 1 | |||||
12 | P | DS-TP x TP-TSred | + | * | + | + | + | 2 | 1 | 1 | ||||||
13 | Q | TS-Ala | + | + | + | + | 986.6 | 986.4 | + | + | + | 1 | ||||
R | HSred-Ac-TP (right lactam reducedh) | + | * | 1720.7 | 1721.6 | |||||||||||
S | HSred-Ac-TP (left lactam reducedh) | + | * | 1720.7 | 1721.7 | |||||||||||
14 | T | DS-TP x TP-TS | + | * | + | + | 2283.3 | 2283.4 | + | + | + | 2 | 1 | 1 | ||
U | HS-TP-Ac | + | * | + | 1734.7 | 1734.6 | + | + | + | 2 | 1 | 1 | ||||
17 | V | TS-TP x TP-TS | + | * | + | + | + | + | + | 2 | 1 | 1 | ||||
W | HS-Ala-Ac | + | * | 1361.8 | 1361.7 | |||||||||||
X | HS-Ala-Ac | + | * | 1361.8 | 1361.7 | |||||||||||
18 | Y | HS-TP | + | * | + | + | + | + | + | 2 | 1 | 1 | ||||
19 | Z | HS-Ala | + | * | + | + | + | + | + | 1 | ||||||
20 | AA | TS-TP x TP-HS | + | * | + | + | + | + | 2 | 1 | 1 | |||||
Muropeptides produced from germinated spore exudate following mutanolysin digestion | ||||||||||||||||
aG1 | TriS-TP Red | + | 1141.5 | 1141.4 | + | + | 2 | 1 | 1 | |||||||
G8f | aG4 | TriS-TP | + | 1155.5 | 1155.4 | + | + | + | 2 | 1 | 1 | |||||
G8Ag | aG5 | TriS-Ala | + | 782.5 | 782.4 | + | + | + | 1 | |||||||
aG8 | PS-TP-Ac | + | 1531.6 | 1531.5 | + | + | + | 2 | 1 | 1 | ||||||
aG9 | PS-Ala-Ac | + | 1160.1 | 1159.9 | + | + | + | 1 | ||||||||
aG10d | TriS-TP x TP-TriS | + | 2293.8 | 2293.8 | + | + | + | 2 | 1 | 1 | ||||||
G8Bg | aG11 | PS-TP | + | 1573.7 | 1573.6 | + | + | + | 2 | 1 | 1 | |||||
Muropeptides produced from germinated spore exudate with no mutanolysin digestion | ||||||||||||||||
G2f | aG2 | TS-TP NAGr Red | + | 1344.8 | 1344.6 | + | + | + | 2 | 1 | 1 | |||||
G1f | aG3 | TS-Ala NAGr Red | + | 972.7 | 972.6 | + | + | + | 2 | 1 | 1 | |||||
G3f | aG6 | TS-TP NAGr | + | 1358.6 | 1358.5 | + | + | + | 2 | 1 | 1 | |||||
G4f | aG7 | TS-Ala NAGr | + | + | 986.6 | 986.4 | + | + | + | 1 | ||||||
aG10u | HS-TP-Ac NAGr | + | 1734.7 | 1735.2 | + | + | + | 2 | 1 | 1 | ||||||
aG12 | HS-Ala-Ac NAGr | + | 1361.8 | 1361.8 | + | + | + | 1 | ||||||||
G6f | aG13 | HS-TP NAGr | + | 1776.8 | 1777.2 | + | + | + | 2 | 1 | 1 |
Numbering as in reference 23.
DS, disaccharide (NAG-NAM); TS, tetrasaccharide (NAG-lactam-NAG-NAM); HS, hexasaccharide (NAG-lactam-NAG-lactam-NAG-NAM); TriS, trisaccharide (lactam-NAG-NAM); PS, pentasaccharide (lactam-NAG-lactam-NAG-NAM); TriP, tripeptide (Ala-Glu-Dpm); TP, tetrapeptide (Ala-Glu-Dpm-Ala); Ac, deacetylated glucosamine; +Am, amidated Dpm; Red, reduced lactam (an artifact of sample preparation [30])); NAGr, NAG at the reducing end; x, cross-link between two peptides.
A plus sign indicates that the muropeptide was detected in the sample, was integrated for structural parameter determinations (Table 2), and is indicated in Fig. 2. An asterisk indicates that the muropeptide was detected in the sample and was integrated for structural parameter determinations (Table 2) but is very small and not indicated in Fig. 2.
m/z expected is the mass-to-charge ratio predicted for the deprotonated ion of the muropeptide structure indicated. m/z observed is the value determined using matrix-assisted laser desorption ionization-time of flight mass spectrometry or electrospray ionization mass spectrometry.
The analysis method used does not produce reliable quantitative results for amino sugars. A plus sign indicates that the compound was clearly detected. For amino acids, the values are the molar ratios, which are rounded to the nearest whole number.
Numbering as in reference 6.
Numbering as in reference 7.
The position of lactam reduction was determined by fragmentation of the muropeptide during tandem mass spectrometry analysis.