Much more sensitive substrate for leukocyte proteinase 3 (PR-3, myeloblastin) (Kcat/km=1.06 x 106 M-1s-1, Km=63 µM) than MeOSuc-AAPV-pNA (Chromogenic Substrate) (Prod. No. BML-P213) (Kcat/km=1.06 x 103 M-1s-1, Km=470 µM). Also cleaved by neutrophil elastase (Kcat/km=9.2 x 106 M-1s-1, Km=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 (
Product Details
| Sequence: | t-butyloxycarbonyl-Ala-Ala-Nva-thiobenzyl ester |
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| Formula: | C23H35N3O5S |
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| MW: | 465.6 |
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| Formulation: | Lyophilized. |
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| Purity: | ≥95% (HPLC) |
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| Appearance: | White to off-white powder. |
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| Solubility: | Soluble in DMSO (1mg/ml). |
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| Shipping: | Shipped on Blue Ice |
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| Long Term Storage: | -20°C |
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| Use/Stability: | Stock solutions in DMSO are stable for up to 3 months when stored at -20°C. |
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| Regulatory Status: | RUO - Research Use Only |
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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;
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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
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