Table 1 Kinetic Parameters Determined for AaCHYMO Using Commercially Available Chymotrypsin- and AaCHYMO-Specifically Designed AMC Fluorescent Substrates Compared to Human and Bovine Chymotrypsin.
From: Biochemical and physiological characterization of Aedes aegypti midgut chymotrypsin
Substrate | AaCHYMO | Human Chymotrypsin | Bovine Chymotrypsin | Human/AaCHYMO | Bovine/AaCHYMO | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
kcat (s-1) | Km (µM)* | kcat/Km (M-1 s-1) | Specific Activity | kcat (s-1) | Km (µM)* | kcat/Km (M-1 s-1) | Specific Activity | kcat (s-1) | Km (µM)* | kcat/Km (M-1 s-1) | Specific Activity | kcat/Km | kcat/Km | |
µmol min-1(mg protein)-1 | µmol min-1(mg protein)-1 | µmol min-1(mg protein)-1 | ratio | ratio | ||||||||||
Commercially Available | ||||||||||||||
Suc-Ala-Ala-Pro-Phe-AMC | 4.81 | 9.72 | 4949578 | 11.0 | 608.7 | 114.2 | 5329831 | 1460.8 | 832 | 18.77 | 44326052 | 1997 | 1.08 | 9.0 |
Suc-Ala-Ala-Phe-AMC | 0.92 | 83.16 | 11087 | 2.10 | 8.38 | 30.0 | 279724 | 20.1 | 100.3 | 48.44 | 2069915 | 240.6 | 25.2 | 186.7 |
MeO-Suc-Arg-Pro-Tyr-AMC | 14.1 | 9.52 | 1481520 | 32.1 | 1597 | 12.3 | 129915921 | 3832 | 3146 | 8.014 | 392563015 | 7550 | 87.7 | 265.0 |
Suc-Leu-Leu-Val-Tyr-AMC | 0.06 | 6.72 | 9410 | 0.14 | 597 | 6.6 | 89814954 | 1432.8 | 580.3 | 10.67 | 54389253 | 1393 | 9545 | 5780 |
Suc-Arg-Pro-Phe-His-Leu-Leu-Val-Tyr-MCA | 0.61 | 1.54 | 394925 | 1.38 | 37.4 | 1.2 | 31562676 | 89.8 | 6.82 | 0.57 | 11959064 | 16.4 | 79.9 | 30.3 |
Suc-Leu-Tyr-AMC | 0.07 | 7.57 | 8712 | 0.15 | 5.16 | 10.6 | 485732 | 12.4 | 8.58 | 16.29 | 526499 | 20.6 | 55.8 | 60.4 |
Suc-Ile-Ile-Trp-MCA | 5.14 | 61.68 | 83279 | 11.7 | 794 | 12.2 | 65188834 | 1905.6 | 7893 | 13.37 | 590376465 | 18944 | 782.8 | 7089 |
Ac-Ala-Asn-Trp-AMC | 2.69 | 33.31 | 80636 | 6.12 | 425 | 25.1 | 16932271 | 1020 | 255.9 | 56.16 | 4556624 | 614.2 | 210.0 | 56.5 |
Z-Leu-Leu-Leu-MCA | 0.03 | 0.13 | 204048 | 0.062 | - | - | - | - | - | - | - | - | - | - |
Ac-Pro-Ala-Leu-AMC | 0.02 | 9.00 | 1818 | 0.037 | - | - | - | - | 0.0061 | 7.63 | 801 | 0.014664 | - | 0.44 |
Z-Gly-Gly-Leu-AMC | - | - | - | - | - | - | - | - | - | - | - | - | - | - |
Z-Val-Lys-Met-MCA | 0.07 | 1.21 | 60149 | 0.166 | 0.41 | 6.4 | 64896 | 0.991 | 6.32 | 6.06 | 1043215 | 15.2 | 1.08 | 17.3 |
Boc-Val-Leu-Lys-AMC | - | - | - | - | 5.17 | 225.5 | 22942 | 12.4 | - | - | - | - | - | - |
Z-Arg-Arg-AMC | - | - | - | - | 4.79 | 568.2 | 8424 | 11.5 | - | - | - | - | - | - |
AaCHYMO-Specifically Designed | ||||||||||||||
Ac-Ala-His-Val-Phe-AMC | 26.9 | 8.11 | 33173056 | 61.3 | 83.8 | 30.0 | 2797242 | 201.2 | 339.7 | 31.0 | 10974690 | 815.2 | 0.084 | 0.331 |
Ac-Ala-His-Leu-Phe-AMC | 10.6 | 4.40 | 2419697 | 24.3 | 23.4 | 13.7 | 1706754 | 56.2 | 40.1 | 14.1 | 2849692 | 96.2 | 0.705 | 1.18 |
Ac-Ala-His-Val-Tyr-AMC | 0.817 | 7.04 | 116115 | 1.86 | 8.4 | 25.9 | 325097 | 20.2 | 8.58 | 23.9 | 359737 | 20.6 | 2.80 | 3.10 |
Ac-Ala-His-Leu-Tyr-AMC | 6.07 | 1.24 | 4888531 | 13.8 | 248.3 | 12.7 | 19548556 | 595.8 | 642.7 | 19.8 | 32408808 | 1542.4 | 4.00 | 6.63 |
Ac-Ala-His-Thr-Trp-AMC | 0.072 | 7.30 | 9815 | 0.163 | 190.0 | 25.1 | 7583500 | 455.9 | 339.7 | 19.9 | 17094447 | 815.2 | 772.6 | 1741.7 |
Ac-Ala-His-Asp-Trp-AMC+ | - | - | - | - | 48.5 | 14.8 | 3285908 | 116.4 | 272.7 | 20.2 | 13525132 | 654.4 | - | - |
Ac-Ala-His-Val-Trp-AMC | 5.51 | 6.32 | 872249 | 12.6 | 438.7 | 18.9 | 23222163 | 1052.8 | 1839.7 | 15.3 | 120318291 | 4415.2 | 26.6 | 137.9 |
Ac-Ala-His-Val-Leu-AMC | 5.42 | 3.95 | 1372350 | 12.3 | 1.26 | 33.1 | 38176 | 3.03 | 2.30 | 31.9 | 72076 | 5.51 | 0.028 | 0.053 |
Ac-Pro-His-Val-Met-AMC | 133.1 | 10.7 | 12482802 | 303.4 | 2.9 | 18.3 | 157534 | 6.90 | 4.19 | 11.3 | 371420 | 10.1 | 0.013 | 0.030 |
Ac-Ala-His-Val-Met-AMC | 2.83 | 4.30 | 658452 | 6.46 | 8.61 | 43.8 | 196589 | 20.7 | 8.93 | 29.7 | 300460 | 21.4 | 0.299 | 0.456 |
