Prothrombin, Fragment 1.2, Human

Prothrombin is a vitamin K-dependent plasma protein which is synthesized in the liver (1). Prior to secretion into plasma, prothrombin undergoes post-translational modification by a vitamin K-dependent carboxylase which converts ten specific glutamic acid residues to g-carboxyglutamic acid (gla). The ten gla residues are located within the first 40 amino acids of the mature protein and contribute to the ability of prothrombin to bind to negatively charged phospholipid membranes. Prothrombin contains two regions of internal homology which are referred to as "kringle" structures. These regions of conspicuous secondary structure are located between residues 40 and 270 of the mature plasma protein and replace the growth factor domains found in several other plasma serine proteases. Thus far, no function has been ascribed to these regions, but there is suspicion that they may play a role in one of several binary protein interactions involving prothrombin. The mature single chain protein circulates in plasma as a zymogen and, during coagulation, is proteolytically activated to the potent serine protease a-thrombin. This proteolysis is catalyzed by the prothrombinase enzyme complex. During activation, human prothrombin is cleaved at Arg273-Thr274 and at Arg323-Ser324 to yield a "pro" fragment (fragment 1.2) and thrombin, the latter of which is composed of two chains covalently linked by a disulfide bond.\n\n\nLocalization: Plasma \n\nMolecular weight: Pre-cleavage Prothrombin 72,000 (1,4,5)\n\nMolecular weight: Prothrombin Fragment 1.2 34,566\n\nPlasma Concentration: 100ug/ml (1)\n\nMode of Action: Zymogen; precursor to the serine protease a-thrombin\n\nExtinction Coefficient: E1%1cm, 280nm=13.8 (4)\n\nIsoelectric Point: 4.7-4.9 (6)\n\nStructure: Single chain, NH2-terminal gla domain, two kringle regions \n\nPercent Carbohydrate: 8.2% (4)\n\nPost-translational Modifications: Ten gla residues (4,5)\n\nStorage and Stability: May be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

P9115-12

Size

1mg

Form

Supplied as a liquid in 50% glycerol/H2O.

Purity

(same/more than)95% by SDS-PAGE. Human prothrombin is prepared from fresh frozen human plasma as described by Bajaj, et al. (2). Purity is determined by SDS-PAGE analysis, and activity is measured by clotting and/or chromogenic substrate assay, following conversion of prothrombin to thrombin.

References

1. Mann, K.G., et al., Methods in Enzymology, 45, 156 (1976). 2. Bajaj, S.P., et al., Prep. Biochem., 11, 397 (1981). 3. Owen, W.G., et al., J. Biol. Chem., 249, 594 (1974). 4. Kisiel, W., et al., Biochem. Biophys. Acta, 304, 103 (1973). 5. Magnusson, S., et al., In Proteases in Biological Control, ed. E. Reich, D.B. Rifkin, E. Shaw, pp. 123-149. New York: Cold Spring Harbor Laboratories, 1975. 6. Discipio, R.G., et al., Biochemistry, 18, 899 (1979).