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EFFECT
OF CTI ON THE APTT AND PT OF NORMAL HUMAN PLASMA. The
graph illustrates the effect of added CTI on the PT and aPTT of normal
human plasma. CTI was added to normal pooled human plasma to various
final concentrations, and the mixtures were allowed to incubate for a
brief period on ice prior to conducting the assays. The PT remains
unaffected while the aPTT doubles with the addition of approximately 60
micrograms of CTI per ml of plasma.
Corn trypsin inhibitor (CTI) is a small protein that is localized in the kernels of most species of corn. CTI is not only an inhibitor of trypsin, but is also a
specific human factor XIIa inhibitor when tested in blood clotting experiments (1-6). The inhibitor forms a one-to-one complex with either trypsin or factor XIIa, and when added to plasma, prolongs the activated partial thromboplastin time without affecting the PT assay (6). The specificity for factor XIIa makes the inhibitor useful for the segregation and study of coagulation reactions (6).
The protein is comprised of 112 amino acids which yields a calculated molecular weight of 12,028 (7). The calculated molecular weight and that which has been determined by sedimentation equilibrium analyses of the purified protein (12,500) are in good agreement (3). A comparison of the amino acid sequence of CTI to that of other trypsin or serine protease inhibitors reveals little or no similarities (7).
CTI is purified using a combination of published procedures (3,4). To begin the purification process, CTI is extracted from the kernels of fresh sweet corn into a physiologic buffer. The extract is then de-fatted using acetone, and the protein is further purified by employing gel filtration and ion-exchange chromatography. The final CTI preparation appears as a single band by SDS-PAGE analyses under both reducing and non-reducing conditions.
Preparations of CTI are tested for the ability to prolong the aPTT assay without affecting the PT assay. The specific activity of each lot of CTI is determined, and one unit is defined as the amount of CTI required to double the aPTT of normal human plasma.
Purified CTI is formulated in 20 mM Tris, 0.15 M NaCl, pH 7.4 and should be stored frozen at -20°C or colder.
Blood
Collection Tubes
To
simplify the process of collecting samples with added CTI, HTI has
developed blood collection tubes containing
CTI. These tubes simplify the process of conducting TF-dependent studies
by allowing you to draw blood directly
onto an anticoagulant containing CTI. You may choose to use our standard
CTI/Citrate formulation (11mM Citrate, 50 ug/mL
CTI) or you may create your own custom formulation. HTI’s blood
collection tubes are not sterile and
are manufactured and sold for research use only.
DOWNLOAD
CORN TRYPSIN INHIBITOR APPLICATION PDF
Properties of
Corn Trypsin Inhibitor
| Localization: |
Corn
Kernals |
| Mode of action: |
forms a
one-to-one complex with either trypsin or factor XIIa
and inhibits their respective catalytic activity. |
| Molecular weight: |
12,500 (3) |
| Extinction coefficient: |
| E |
|
= 20.0
(determined by ultra centrifugation studies) |
|
| Structure: |
single
chain protein comprised of 112 amino acids (3,7) |
|
Catalog
Number |
Description |
|
CTI-01 |
Corn Trypsin Inhibitor |
|
SCAT-27-4.5/5 |
CTI/Citrate Blood
Collection Tube |
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References
1. Hochstrasser, K., et al., Z. Physiol. Chem., 348, 1337 (1967).
2. Halim, R.K., et al., Crop. Sci., 13, 405 (1973).
3. Swartz, M.J., et al., J. Biol. Chem., 252, 8105 (1977).
4. Hojima, Y., et al., Thromb. Res., 20, 149 (1980).
5. Ratnoff, O.D. and Moneme, V., Proc. Soc. Exp. Biol. Med., 166, 297 (1981).
6. Rand, M.D. et al., Blood, 88, 3432 (1996).
7. Mahoney, W.C., et al., J. Biol. Chem., 259, 8412 (1984).
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