202
In summary, the LC-MS/MS analysis of both tryptic and GluC V8 digests
allowed the identification of a total of 30 glycation sites on the lysine residues
(Fig. 11.9a). Mapping these glycation sites on the 3D representation of the BSA
(Fig. 11.9b, lysines highlighted in red) permitted to observe that they correspond to
lysine residues located at the outer surface of the BSA. In addition, the number of
Table 11.3 Glycopeptides identified in the bovine serum albumin protein by LC-ESI-QqTOF-MS/
MS analysis of the hapten-BSA glycoconjugate digested with the endoproteinase GluC V8 [69]
Precursor ion
m/z
Deviation Missed
(charge)
Mr (expt) Mr (calc) Da
Cleavage Peptide
698.3598
(+2)
1394.7051 1394.6803 0.0248
0
K
* LGE (Lys 420)
790.3774
(+2)
1578.7403 1578.7288 0.0115
1
K
* QEPE (Lys ll7)
844.3721
(+2)
1686.7297 1686.7499 −0.0202 1
EFK
* ADE (Lys 151)
894.4637
(+2)
1786.9128 1786.8976 0.0152
0
HVK
*
LVNE (Lys 65)
862.9539
(+2)
1723.8933 1723.8754 0.01179 1
VTK
* LVTD (Lys 256)
897.4200
(+2)
1792.8255 1792.8176 0.0079
0
K
* SHCIAE (Lys 309)
902.4801
(+2)
1802.9456 1802.9289 0.0167
0
LTKVHK
* E (Lys 266)
962.1552
(+2)
2883.4439 2883.4093 0.0346
0
VSRSLGK
* VGTRCCTKPE (Lys
455)
987.9658
(+2)
1973.9171 1973.8949 0.0222
0
K
* VTKCCTE (Lys 495)
989.9697
(+2)
1977.9248 1977.9154 0.0094
2
K
* QEPERNE (Lys 117)
992.4568
(+2)
1982.8989 1982.884 0.0149
1
CCDK
*
PLLE (Lys 304)
995.4702
(+2)
1988.9258 1988.8912 0.0346
1
FAK
*
TCVADE (Lys 75)
1013.0142
(+2)
2024.0138 2024.0123 0.0015
0
LCK
* VASLRE (Lys 100)
1048.5225
(+2)
2095.0305 2094.9984 0.0321
1
K
* SLHTLFGDE (Lys 88)
1097.0595
(+2)
2192.1045 2192.0909 0.0136
0
DK
*
GACLLPKIE (Lys 197)
1124.5018
(+2)
2246.9890 2246.9876 0.0014
1
DK
*
DVCKNYQE (Lys 336)
1336.1446
(+2)
2670.2746 2670.2728 0.0018
0
LLYYANK
* YNGVFQE (Lys 183)
1156.132
(+2)
2310.2495 2310.2345 0.0150
0
YAVSVLLRLAK
*
E (Lys 374)
M. Bologna et al.
In summary, the LC-MS/MS analysis of both tryptic and GluC V8 digests
allowed the identification of a total of 30 glycation sites on the lysine residues
(Fig. 11.9a). Mapping these glycation sites on the 3D representation of the BSA
(Fig. 11.9b, lysines highlighted in red) permitted to observe that they correspond to
lysine residues located at the outer surface of the BSA. In addition, the number of
Table 11.3 Glycopeptides identified in the bovine serum albumin protein by LC-ESI-QqTOF-MS/
MS analysis of the hapten-BSA glycoconjugate digested with the endoproteinase GluC V8 [69]
Precursor ion
m/z
Deviation Missed
(charge)
Mr (expt) Mr (calc) Da
Cleavage Peptide
698.3598
(+2)
1394.7051 1394.6803 0.0248
0
K
* LGE (Lys 420)
790.3774
(+2)
1578.7403 1578.7288 0.0115
1
K
* QEPE (Lys ll7)
844.3721
(+2)
1686.7297 1686.7499 −0.0202 1
EFK
* ADE (Lys 151)
894.4637
(+2)
1786.9128 1786.8976 0.0152
0
HVK
*
LVNE (Lys 65)
862.9539
(+2)
1723.8933 1723.8754 0.01179 1
VTK
* LVTD (Lys 256)
897.4200
(+2)
1792.8255 1792.8176 0.0079
0
K
* SHCIAE (Lys 309)
902.4801
(+2)
1802.9456 1802.9289 0.0167
0
LTKVHK
* E (Lys 266)
962.1552
(+2)
2883.4439 2883.4093 0.0346
0
VSRSLGK
* VGTRCCTKPE (Lys
455)
987.9658
(+2)
1973.9171 1973.8949 0.0222
0
K
* VTKCCTE (Lys 495)
989.9697
(+2)
1977.9248 1977.9154 0.0094
2
K
* QEPERNE (Lys 117)
992.4568
(+2)
1982.8989 1982.884 0.0149
1
CCDK
*
PLLE (Lys 304)
995.4702
(+2)
1988.9258 1988.8912 0.0346
1
FAK
*
TCVADE (Lys 75)
1013.0142
(+2)
2024.0138 2024.0123 0.0015
0
LCK
* VASLRE (Lys 100)
1048.5225
(+2)
2095.0305 2094.9984 0.0321
1
K
* SLHTLFGDE (Lys 88)
1097.0595
(+2)
2192.1045 2192.0909 0.0136
0
DK
*
GACLLPKIE (Lys 197)
1124.5018
(+2)
2246.9890 2246.9876 0.0014
1
DK
*
DVCKNYQE (Lys 336)
1336.1446
(+2)
2670.2746 2670.2728 0.0018
0
LLYYANK
* YNGVFQE (Lys 183)
1156.132
(+2)
2310.2495 2310.2345 0.0150
0
YAVSVLLRLAK
*
E (Lys 374)
M. Bologna et al.