Multiple Active Forms of Thrombin

• Published in: The Journal of biological chemistry Vol. 248; no. 5; pp. 1868 - 1875
• Main Authors: Mann, Kenneth G., Yip, Ray, Heldebrant, Charles M., Fass, David N.
• Format: Journal Article
• Language: English
• Published: American Society for Biochemistry and Molecular Biology 10-03-1973
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)44270-1

The polypeptide chain location of active site serine and carbohydrate in the multiple forms of thrombin present in Parke-Davis
thrombin have been investigated by means of [ 3 H]diisopropylfluorophosphate incorporation and carbohydrate staining techniques. Both the sodium dodecyl sulfate electrophoretic
technique and gel filtration in 6 m guanidinium chloride have been applied to the resolution of individual thrombin forms and their component chains.
The results of these studies indicate that the active site serine is contained in the 33,000-dalton chain of the large (39,000
dalton) thrombin, and in the 18,000- and 14,000-dalton chains of the smaller (28,000 dalton) thrombins. Carbohydrate analysis
of the resolved thrombin components indicate that, although the large thrombin contains covalently attached carbohydrate,
the smaller enzymes do not.