Kinetics of Thrombin Inactivation as Influenced by Physical Conditions, Trypsin, and Serum

Glazko, Anthony J.; Ferguson, John H.

doi:10.1085/jgp.24.2.169

Cited by 10 publications

(1 citation statement)

References 21 publications

(28 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Most of the experiments described here involve changes in thrombin concentration which can be measured indirectly by means of the clotting time (18). In all cases, solutions were maintained at 380 C., and the pH was kept in the range of 7.2 to 7.6 as measured with a glass electrode.…”

Section: Methodsmentioning

confidence: 99%

Effect of Blood Protease and Trypsin Inhibitor on the Clotting Mechanism 1

Glazko¹

1947

J. Clin. Invest.

Self Cite

View full text Add to dashboard Cite

The influence of proteolytic enzymes on blood coagulation has attracted considerable attention in recent years because of the possibility of their involvement in the normal clotting process. Eagle and Harris (1) showed that certain proteolytic enzymes such as trypsin could activate prothrombin; while other enzymes such as papain could clot fibrinogen directly. Schmitz (2, 3, 4) claimed that he isolated both trypsin and a specific trypsin-inhibitor from plasma. However, more recent work indicates that the protease is not identical with trypsin, although it is quite similar in many respects (5,6).It has long been known that proteolytic activity appears in blood following treatment with chloroform, acetone, or certain other denaturing agents. Tagnon and others (7,8,9) used this technic to activate blood protease and described many of its properties. Holmberg (10) and Christensen (11) independently demonstrated that streptokinase,2 which is itself devoid of proteolytic activity, could activate the protease in blood and thus produce digestion. Christensen also showed that the blood protease activated by streptokinase was identical with chloroform-activated serum protease (6). The inactive enzyme is located principally in globulin fraction III-2, along with considerable amounts of prothrombin (12, 13). Active protease is present in globulin fraction III-3, which is actually a sub-fraction of III-2. Fraction III-3 was used in the present experiments as a convenient source of blood protease.A definite coagulation-inhibiting effect of crystalline pancreatic trypsin-inhibitor was demon-1 This work was supported in part by grants from the Life Insurance Medical Research Fund and the University Center in Georgia.2 Following the suggestion of Christensen (11), it was decided to adopt the term "streptokinase" for use throughout this paper. The earlier designation "streptococcal fibrinolysin" was based on the erroneous assumption that the fibrinolytic enzyme itself was produced in the bacterial culture. strated by Ferguson (14), supporting his thesis that a protease is involved in the activation of prothrombin (15). Similar results were obtained by Grob (16), who also found that serum anti-protease prepared by the method of Schmitz (4) delayed coagulation. Tagnon and Soulier (17) showed that trypsin-inhibitor isolated from soy bean flour could also inhibit coagulation. Their work has been confirmed and extended by the observations described in this paper. METHODS AND MATERIALSMost of the experiments described here involve changes in thrombin concentration which can be measured indirectly by means of the clotting time (18). In all cases, solutions were maintained at 380 C., and the pH was kept in the range of 7.2 to 7.6 as measured with a glass electrode. A small amount of phenyl mercuric borate was added to the reagents to prevent bacterial growth. Tests for thrombic activity were carried out by pipetting 0.5 ml. of thrombin into 1 ml. of fibrinogen solution, using 10 mm. X 75 mm. serological tubes. Clot formation was timed fr...

show abstract

Section: Methodsmentioning

confidence: 99%

Effect of Blood Protease and Trypsin Inhibitor on the Clotting Mechanism 1

Glazko¹

1947

J. Clin. Invest.

Self Cite

View full text Add to dashboard Cite

show abstract

Blood Clotting and Related Processes

Astrup¹

1950

Advances in Enzymology - And Related Areas of Molecular Biology

View full text Add to dashboard Cite

Protoplasmic clotting in isolated muscle fibers

Woodward

1948

J. Cell. Comp. Physiol.

View full text Add to dashboard Cite

FOUR FIGURESChanges in the colloidal state of protoplasm provide an important index to the nature of intracellular events. Certain of these changes appear to be fundanlentally identical with the surface precipitation reaction (s.p.r.) which occurs ~7lle11 the membrane of a cell is ruptured, and the cellular contents are exposed to the external medium. The capacity to show this reactioii seeins to be a general property of protoplasm, and it has been demonstrated, under the proper conditions, in many kinds of cells. A substantial amount of evidence suggests that it is the result of the same kind of process as that which causes the gelation of the interior of a cell following stimulation and, also, that it iiivolves a mechanism similar to that operating in the coagulation of blood and other kinds of biological fluids, such as milk. ( P o r a more detailed description and discussion of the s.p.r. and references to experiments on it, see Heilbrunn, '43.)Usually tlie s.p.r. is produced by crushing or tearing a cell. These methods unfortunately tend to permit the use of the reaction only as a qualitative test; that is, it either o c c~~r s o r does not occur in the presence of a given medium. Evidence concerning the degree of effect under given experimental . A dissertation submittcd to the facultp of the Dqxirtment of Zoologp of the Univcrsity of Pennsylvania in partial fulfillment of the requirements for the degree of Doctor of Philosophy. The research was supported in part by :I grant from the Roekefeller Youudatioii administcred by Dr. L. V. Heilbrunn. 359

show abstract

Kinetics of Thrombin Inactivation as Influenced by Physical Conditions, Trypsin, and Serum

Cited by 10 publications

References 21 publications

Effect of Blood Protease and Trypsin Inhibitor on the Clotting Mechanism 1

Effect of Blood Protease and Trypsin Inhibitor on the Clotting Mechanism 1

Blood Clotting and Related Processes

Protoplasmic clotting in isolated muscle fibers

Contact Info

Product

Resources

About