Much more sensitive substrate for leukocyte proteinase 3 (PR-3, myeloblastin) (K
cat/k
m=1.06 x 10
6 M
-1s
-1, K
m=63 µM) than MeOSuc-AAPV-pNA (Chromogenic Substrate) (Prod. No.
BML-P213) (K
cat/k
m=1.06 x 10
3 M
-1s
-1, K
m=470 µM). Also cleaved by neutrophil elastase (K
cat/k
m=9.2 x 10
6 M
-1s
-1, K
m=2.2 µM), granzyme H, cathepsin G, chymotrypsin, and rat MCPI and II.Typical assay conditions: substrate concentration 88-250 µM; enzyme concentration 0.2-20 nM; buffer [100 mM HEPES pH 7.5, 500 mM NaCl, 10% DMSO]; with 170 µM either 5,5’-dithiobis(2-nitrobenzoic acid) (DTNB) or 4,4’-dithiobis(pyridine) (from 15 mM DMSO stocks); 25°C.
Hydrolysis of substrate can be detected at 412 nm for DTNB (ε
405=13,600) or 324 nm for 4,4’-dithiobis(pyridine) (ε
405=19,800).
Product Specification
| Formula: | C23H35N3O5S |
| |
| MW: | 465.2 |
| |
| Purity: | ≥95% (HPLC) |
| |
| Appearance: | White to off-white powder. |
| |
| Sequence: | t-butyloxycarbonyl-Ala-Ala-Nva-thiobenzyl ester |
| |
| Peptide Content: | 60-80% |
| |
| Solubility: | Soluble in methanol or DMSO (≥15mM). |
| |
| Long Term Storage: | -20°C |
| |
| Use/Stability: | Stock solutions in DMSO are stable for up to 3 months when stored at -20°C. |
| |
Product Literature References
The human cytotoxic T cell granule serine protease granzyme H has chymotrypsin-like (chymase) activity and is taken up into cytoplasmic vesicles reminiscent of granzyme B-containing endosomes: K.M. Edwards, et al.; J. Biol. Chem.
274, 30468 (1999),
Abstract;
Full Text
Proteinase 3: substrate specificity and possible pathogenetic effect of Wegener's granulomatosis autoantibodies (c-ANCA) by dysregulation of the enzyme: K.M. Dolman, et al.; Adv. Exp. Med. Biol.
336, 55 (1993),
Abstract;
Substrate and inhibitor studies on proteinase 3: C.M. Kam, et al.; FEBS Lett.
297, 119 (1992),
Abstract;
Active site mapping of the serine proteases human leukocyte elastase, cathepsin G, porcine pancreatic elastase, rat mast cell proteases I and II. Bovine chymotrypsin A alpha, and Staphylococcus aureus protease V-8 using tripeptide thiobenzyl ester substra: J.W. Harper, et al.; Biochemistry
23, 2995 (1984),
Abstract;
General Literature References
Compared action of neutrophil proteinase 3 and elastase on model substrates. Favorable effect of S'-P' interactions on proteinase 3 catalysts: C. Koehl, et al.; J. Biol. Chem.
278, 12609 (2003),
Abstract;
Full Text