Table 1.
Percent hydrolysis of pro-KLK(1–15) fusion proteins for 100:1 molar ratio incubation with mature KLK9, 10, and 15 at pH 6.0/7.4 for 1 h and 24 h at 37°C.
| P1 | KLK9 | KLK10 | KLK15 | ||||
|---|---|---|---|---|---|---|---|
| 1 h | 24 h | 1 h | 24 h | 1 h | 24 h | ||
| pro-KLK1 | Arg | – | – | – | 0/5 | 1/9 | 34/62 |
| pro-KLK2 | Arg | – | 5/13 | – | 1/9 | 1/11 | 34/72 |
| pro-KLK3 | Arg | – | 4/8 | – | 8/3 | 0/8 | 32/77 |
| pro-KLK4 | Gln | – | – | – | – | – | – |
| pro-KLK5 | Arg | – | – | – | – | 0/4 | 16/47 |
| pro-KLK6 | Lys | – | – | – | – | – | – |
| pro-KLK7 | Lys | – | – | – | 3/14 | 3/10 | 39/77 |
| pro-KLK8 | Lys | – | – | – | 2/11 | 3/16 | 43/86 |
| pro-KLK9 | Arg | – | –/14 | – | 0/7 | 3/10 | 27/64 |
| pro-KLK10 | Arg | – | – | – | 0/6 | 0/0 | 3/24 |
| pro-KLK11 | Arg | – | –/20 | – | – | 0/4 | 18/71 |
| pro-KLK12 | Lys | – | – | – | – | – | 1/12 |
| pro-KLK13 | Lys | – | –/21 | – | – | 0/2 | 10/30 |
| pro-KLK14 | Lys | – | – | – | 0/7 | 0/5 | 14/54 |
| pro-KLK15 | Lys | – | – | – | 2/10 | 6/16 | 46/91 |
Mature KLK10 and KLK15 proteins were expressed from an Escherichia coli host, refolded and purified as previously described (Debela et al., 2006). Mature KLK9 was expressed as recombinant protein from HEK293 human embryonic kidney epithelial cells as the expression host, as previously described for pro-KLK6 (Yoon et al., 2008). The pro-KLK9 was activated to mature KLK9 by incubation with enterokinase, as described previously for activation of pro-KLK6 (Blaber et al., 2007). Assay conditions utilized 40 mm of pro-KLK fusion proteins as previously described (Yoon et al., 2007).