Supplemental Top-down mass spectrometry of intact human salivary Cystatins S, S1, and S2. Parent molecular ions were subjected to CAD on a hybrid linear ion-trap FTICR mass spectrometer (A, D, and G). Amino acid sequence and protein sequence coverage is displayed with the graphical output of ProSight PC 2.0 to illustrate assigned ions from the CAD data set (B, E, and H). The location of b and y fragment ions are shown in blue, and highlighted in orange are cysteine residues oxidized to disulfides (-1.00782509 Da each) and phosphorylation of Ser21 and Ser23 (+79.966333 Da). The raw data file and .PUF file for each data set are shown in (C), (F), and (I). (A) (B) P-Score: 1.71E-57 Matching fragments: 44 Non-matching fragments: 63 Fragments explained: 41% Cystatin S 14175.8569 Da (C) Cystatin S CAD.ra cr032709_from030909_crWS_fraction55_ii1182.9962_AMW14183.87 #1 RT:1.47 AV:1 N L:6.41E 4 T:FTMS + p NSIFullms2 1184.00@ cid0.00 [325.00-2000.00] 1174 1175 1176 1177 1178 1179 1180 1181 1182 1183 1184 1185 1186 1187 1188 1189 1190 1191 1192 1193 1194 m/z 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 1183.16 1182.83 1183.33 1183.50 1182.58 1183.66 1184.41 1184.83 1181.66 1182.25 1185.08 1180.98 1186.06 1180.04 1188.28 1176.52 1187.44 1189.13 1177.74 1178.38 1175.33 1191.37 1189.85 1193.17 1174.67 1194.5 cr032709_from 030909_crW S_fraction55_C AD 1182.9962 #1 R T:5.35 AV:1 N L:6.91E3 T:FTMS + p NSI Full m s2 1184.00@ cid10.00 [325.00-2000.00] 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 m/z 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 1184.41 1231.70 1560.94 1343.65 1362.04 1283.42 910.61 1144.60 1111.27 1413.80 835.41 967.47 1087.51 1470.11 1741.24 879.44 1630.76 572.80 1861.30 824.91 1546.72 440.44 648.28 1973.14 735.39 494.86 381.16 Cystatin S.p
Transcript
Supplemental Top-down mass spectrometry of intact human salivary Cystatins S, S1, and S2. Parent molecular ions were subjected to CAD on a hybrid linear ion-trap FTICR mass spectrometer (A, D, and G). Amino acid sequence and protein sequence coverage is displayed with the graphical output of ProSight PC 2.0 to illustrate assigned ions from the CAD data set (B, E, and H). The location of b and y fragment ions are shown in blue, and highlighted in orange are cysteine residues oxidized to disulfides (-1.00782509 Da each) and phosphorylation of Ser21 and Ser23 (+79.966333 Da). The raw data file and .PUF file for each data set are shown in (C), (F), and (I).
Top-down mass spectrometry of intact Cystatin SA, mass 14336.9856 Da. A parent molecular ion (inset) was subjected to CAD on a hybrid linear ion-trap FTICR mass spectrometer (A) . Amino acid sequence and protein sequence coverage is displayed with the graphical output of ProSight PC 2.0 to illustrate assigned ions from the CAD experiment (B) . The location of b and y fragment ions are shown in blue, and highlighted in orange are cysteine residues oxidized to disulfides (-1.00782509 Da each). The raw data file and .PUF file for the data set are shown in (C).
Top-down mass spectrometry of intact Cystatin SN, mass 14303.1553 Da. (A) A parent molecular ion (inset) was subjected to CAD on a hybrid linear ion-trap FTICR mass spectrometer. (B) Amino acid and protein sequence coverage is displayed with the graphical output of ProSight PC 2.0 to illustrate assigned ions from the CAD experiment. The location of b and y fragment ions are shown in blue, and highlighted in orange are cysteine residues oxidized to disulfides (-1.00782509 Da each). The raw data file and .PUF file for the data set are shown in (C).
Top-down mass spectrometry of intact Cystatin C, mass 13334.5829 Da. (A) A parent molecular ion (inset) was subjected to CAD on a hybrid linear ion-trap FTICR mass spectrometer. (B) Amino acid and protein sequence coverage is displayed with the graphical output of ProSight PC 2.0 to illustrate assigned ions from the CAD experiment. The location of b and y fragment ions are shown in blue, and highlighted in orange are cysteine residues oxidized to disulfides (-1.00782509 Da each). The raw data file and .PUF file for the data set are shown in (C).
Cystatin C CAD.raw Cystatin C.puf
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Top-down mass spectrometry of truncated forms of intact Cystatin D with a C46R mutation, with masses 13154.4675 Da and 13596.7015 Da respectively. (A and D) Parent molecular ions (insets) were subjected to CAD on a hybrid linear ion-trap FTICR mass spectrometer. Amino acid and protein sequence coverage is displayed with the graphical output of ProSight PC 2.0 to illustrate assigned ions from the CAD experiments (B and E) . The location of b and y fragment ions are shown in blue, the C46R SNP is reflected in the sequence (SVQR), and highlighted in orange are cysteine residues oxidized to disulfides (-1.00782509 Da each). The raw data file for the CAD data set is shown in (C) and (F).
cr042909_fromcr031009_SM_fraction59_CAD1047.5262 #1 RT: 17.05 AV: 1 NL: 3.26E3T: FTMS + p NSI Full ms2 [email protected] [285.00-2000.00]
