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Enzyme Explorer
Aprotinin
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Physical Properties
While aprotinin and bovine pancreatic trypsin inhibitor (BPTI) are the same protein sequence, the term aprotinin is typically used when describing the protein derived from bovine lung.
Aprotinin is a single peptide chain with three disulfide bonds. Molecular Weight: ~ 65111
E1%280 nm=8.3(water)
pI = 10.56
Inhibitor Activity
Aprotinin is a competitive serine protease inhibitor which forms stable complexes with and blocks the active sites of enzymes. The binding is reversible, and most aprotinin-protease complexes dissociate at pH >10 or <3.2.
| Enzyme |
Inhibition |
|---|
| Acrosin |
Weak inhibition6 |
| Chymotrypsin |
Ki = 9 nM9 |
| Chymotrypsinogen (bovine), pH 8.0 |
Ki = 9 nM4 |
| CMP-Sialic Acid: Lactosylceramide -(2,3)-Sialyltransferase |
74% Inhibition at 300 nM9 |
| Elastase (human leukocytes), pH 8.0 |
Ki = 3.5 µM4 |
| Kallikrein (pancreatic), pH 8.0 |
Ki = 1.0 nM4 |
| Kallikrein (plasma) |
Ki = 30 nM; 100 nM9 |
| Kallikrein (tissue) |
Ki = 1 nM9 |
| Kallikrein (urine) |
Ki = 1.7 nM9 |
| Plasmin (porcine), pH 7.8 |
Ki = 4.0 nM4 |
| Plasminogen activator |
Ki = 8 µM; 27 µM9 |
| Trypsin (bovine), pH 8.0 |
Ki = 0.06 pM4 |
| Trypsinogen (bovine), pH 8.0 |
Ki = 1.8 µM4 |
| Tryptase TL-2 |
16% Inhibition at 10 µM9 |
| Urokinase (human), pH 8.8 |
Ki = 8.0 µM4 |
Unit Definition
One Trypsin Inhibitor Unit (TIU) will decrease the activity of 2 trypsin units by 50%, where 1 trypsin unit will hydrolyze 1.0 µmole of N-α-benzoyl-DL-arginine p-nitroanilide (BAPNA) per minute at pH 7.8 and 25 °C.
Another commonly used unit of activity is the KIU (Kallikrein Inhibitor Unit).
From our data, a conversion factor for Aprotinin is: 1 TIU equals ~1,300 KIU. A published ratio is: 1 TIU equals ~1,025 KIU.10
Solubility and Stability
Aprotinin is freely soluble in water (>10 mg/mL) and in aqueous buffers of low ionic strengths. Dilute solutions are generally less stable than concentrated ones. Solution stability also depends on pH; values of 1-12 can be tolerated. Repeated freeze-thaw cycles should be avoided. The Cys14-Cys38 disulfide bridge is readily split by reducing agents like 2-mercaptoethanol. Due to its compact tertiary structure, aprotinin is relatively stable against denaturation due to high temperature, acids, alkalies, organic solvents or proteolytic degradation (only thermolysin has been found capable of effectively degrading aprotinin after heating to 60-80 °C). The high basicity of aprotinin causes it to adhere to commonly used dialysis tubing and even gel filtration matrices, but the use of acetylated materials and concentrated salt solutions (e.g., 0.1 M NaCl in buffer)3 minimizes the problem. Sterilization may be achieved by filtration through a 0.2 µm filter.
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| Solvent |
Concentration |
Storage Temp. |
% Loss/TIme |
|---|
| Sterile water with 0.9% NaCl and 0.9% benzyl alcohol, pH 5.7-6.2 |
10 mg/mL |
0-5°C |
<4.3%/year5 |
| 2.5% Trichloroacetic acid |
N/A |
80°C |
No loss2 |
| pH <12.6 |
N/A |
N/A |
No loss observed after 24 hrs.7 |
| pH >12 |
N/A |
N/A |
Irreversibly denatured8 |
| pH 7-8 |
0.065-1.95 µg/mL |
4°C |
About 1 week6 |
| pH 7-8 |
0.065-1.95 µg/mL |
-20°C |
>6 months6 |
Related Products
| Product No. |
Description |
|---|
| A6106 |
Aprotinin from bovine lung, BioUltra, ≥4 TIU/mg solid, ≥98% |
| A6103 |
Aprotinin from bovine lung, BioUltra, recombinant, expressed in Nicotiana, ≥5 TIU/mg protein, ≥98% |
| A6279 |
Aprotinin from bovine lung, Saline solution, 3-7 TIU/mg protein |
| A1153 |
Aprotinin from bovine lung, lyophilized powder, 3-8 TIU/mg solid |
| A4529 |
Aprotinin from bovine lung, lyophilized powder, 3-7 TIU/mg solid |
| A3428 |
Aprotinin from bovine lung, lyophilized powder, 3-8 TIU/mg solid, cell culture tested |
| A5421 |
Aprotinin, immobilized on Eupergit® C |
| A2268 |
Aprotinin-Agaros |
| AP5 |
Aprotinin-Agarose prepacked column 2.5 mL |
| P1860 |
Protease Inhibitor Cocktail for tissue culture media |
| P2714 |
Protease Inhibitor Cocktail for general Use |
| S8830 |
SIGMAFAST™ Protease Inhibitor Cocktail Tablets, EDTA-Free |
| S8820 |
SIGMAFAST Protease Inhibitor Cocktail Tablets for general Use |
References
- Merck Index, 12th Ed., S. Budavari, Ed., # 796, p. 128 (1996).
- J. Gen. Physiol., 19, 991 (1936).
- Hoppe-Seyler's Z. Physiol. Chem., 192, 1 (1930).
- Drug Res., 33(1), No. 4, 479 (1983).
- Our internal data.
- Biochemica Information, 1st Ed., J. Keesey, Ed., Boehringer Mannheim Biochemicals, p. 111, Indianapolis (1987).
- Biochemistry, 7, 3634 (1968).
- Life Sci., 28, 1861 (1981).
- Handbook of Enzyme Inhibitors, 2nd Ed., Part B, H. Zollner, Ed., p. 572, VCH Verlagesgesellschaft, Weinheim (1993).
- Biotechnology, p. 565 (June 1990).
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