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Table 2 HPLC-ESI-LTQ-OT-MS/MS analysis of glycosylated T7Muc10

From: High level in vivo mucin-type glycosylation in Escherichia coli

Peptide description

Peptide sequence

Retention time [min]

m/z measured

[M + H]+theor

Charge state

Δm [ppm]

GalNAc moieties

Relative abundance [%]

1

YTMAPTT

16.93

784.355

784.355

+ 1

− 0.98

0

88.21

18.54

987.434

987.434

+ 1

0.61

1

11.80

2/3/4/7/8

DSTTPAPTTKPTT

11.65

1317.653

1317.653

+ 1

0.62

0

27.64

10.68

1520.733

1520.733

+ 1

− 0.40

1

40.35

9.14

1723.812

1723.812

+ 1

0.31

2

25.10

10.83

1723.812

1723.812

+ 1

− 0.32

2

4.34

10.47

1926.891

1926.891

+ 1

− 0.18

3

2.22

10.03

1065.489

2129.971

+ 2

− 0.37

4

0.36

5/6

DSTTPAPTNKPTT

10.42

1330.649

1330.649

+ 1

0.23

0

21.00

9.82

1533.728

1533.728

+ 1

0.39

1

36.34

8.38

1736.807

1736.807

+ 1

0.13

2

27.71

9.98

1736.807

1736.807

+ 1

− 0.07

2

12.00

8.00

1939.887

1939.887

+ 1

− 0.01

3

2.45

9.93

1939.887

1939.887

+ 1

− 0.96

3

0.43

9.47

1071.987

2142.966

+ 2

− 0.14

4

0.07

9

DSTTPAPTTKNNNNNNNNGGLVPRGSGGGSGHHHHHHHHHH

n.d.

 

4339.9583

 

–

0

0

  1. Proteolytic (AspN) peptides of glycosylated T7MuC10 were identified via HPLC-ESI-LTQ-OT-MS/MS analysis. The theoretical monoisotopic mass increases by 203.07937 u per attached O-acetylgalactosamine (C8O5NH13) moiety. Relative quantification of peptides was conducted using the MassMap® software. Extracted ion chromatograms (XIC) of MS signals were generated using a mass tolerance of 5 ppm. Relative peptide abundances of glycosylated and non-glycosylated peptides were calculated from the respective peptide peak areas. The C-terminal peptide was not detected (n.d.)