PAD incubation of CXC and CC chemokines
| Chemokine . | NH2-terminal sequencea . | Enzyme . | E/Sb . | Incubation timec . | Percentage first Citd . | Percentage second Citd . | Mr shifte . |
|---|---|---|---|---|---|---|---|
| CXCL5 | AGPAAAVLRELRCVC- | rabbit PAD | 1/20 | 90 | 89.8 | 0 | + 1.0 |
| hu PAD2 | 1/200 | 120 | 14.0 | 0 | + 0.4 | ||
| hu PAD4 | 1/200 | 120 | 35.1 | 0 | + 0.6 | ||
| CXCL8 | AVLPRSAKELRCQC- | rabbit PAD | 1/20 | 90 | 100 | 0 | + 1.1 |
| hu PAD2 | 1/200 | 120 | 88.0 | 0 | ND | ||
| hu PAD4 | 1/200 | 120 | 94.5 | 0 | + 1.0 | ||
| CCL17 | ARGTNVGRECC- | hu PAD2 | 1/200 | 120 | 24.8 | 22.6 | + 0.8 |
| hu PAD4 | 1/200 | 120 | 92.1 | 54.3 | + 2.4 | ||
| CCL26 | TRGSDISKTCC- | hu PAD | 1/200 | 120 | 17.1 | ND | + 0.2 |
| hu PAD4 | 1/200 | 120 | 17.8 | ND | + 0.2 | ||
| hu PAD4 | 1/40 | 120 | 100 | ND | + 1.3 |
| Chemokine . | NH2-terminal sequencea . | Enzyme . | E/Sb . | Incubation timec . | Percentage first Citd . | Percentage second Citd . | Mr shifte . |
|---|---|---|---|---|---|---|---|
| CXCL5 | AGPAAAVLRELRCVC- | rabbit PAD | 1/20 | 90 | 89.8 | 0 | + 1.0 |
| hu PAD2 | 1/200 | 120 | 14.0 | 0 | + 0.4 | ||
| hu PAD4 | 1/200 | 120 | 35.1 | 0 | + 0.6 | ||
| CXCL8 | AVLPRSAKELRCQC- | rabbit PAD | 1/20 | 90 | 100 | 0 | + 1.1 |
| hu PAD2 | 1/200 | 120 | 88.0 | 0 | ND | ||
| hu PAD4 | 1/200 | 120 | 94.5 | 0 | + 1.0 | ||
| CCL17 | ARGTNVGRECC- | hu PAD2 | 1/200 | 120 | 24.8 | 22.6 | + 0.8 |
| hu PAD4 | 1/200 | 120 | 92.1 | 54.3 | + 2.4 | ||
| CCL26 | TRGSDISKTCC- | hu PAD | 1/200 | 120 | 17.1 | ND | + 0.2 |
| hu PAD4 | 1/200 | 120 | 17.8 | ND | + 0.2 | ||
| hu PAD4 | 1/40 | 120 | 100 | ND | + 1.3 |
NH2-terminal Arg residues are indicated in bold, cysteine motifs are underlined.
Molar Enzyme/Substrate ratio.
Incubation time in minutes.
Percentages of conversion of NH2-terminal Arg residues to Cit were determined by Edman degradation.
Shift compared to the theoretical average Mr of the uncitrullinated chemokine as determined by ion trap mass spectrometry.