Preparation and Properties of Human Prothrombin Complex

1970 ◽  
Vol 24 (03/04) ◽  
pp. 325-333 ◽  
Author(s):  
G. H Tishkoff ◽  
L. C Williams ◽  
D. M Brown
Keyword(s):  
Molecular Weight ◽  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Specific Activity ◽  
Crystalline Form ◽  
Sedimentation Equilibrium ◽  
Crystalline Complex ◽  
Interaction Product ◽  
Proteolytic Enzyme Inhibitor ◽  
Purification Scheme

SummaryAs a corollary to our previous studies with bovine prothrombin, we have initiated a study of human prothrombin complex. This product has been isolated in crystalline form as a barium glycoprotein interaction product. Product yields were reduced compared to bovine product due to the increased solubility of the barium glycoprotein interaction product. On occasion the crystalline complex exhibited good yields. The specific activity of the crystalline complex was 1851 Iowa u/mg. Further purification of human prothrombin complex was made by removal of barium and by chromatography on Sephadex G-100 gels. The final product evidenced multiple procoagulant activities (II, VII, IX and X). The monomeric molecular weight determined by sedimentation equilibrium in a solvent of 6 M guanidine-HCl and 0.5% mercaptoethanol was 70,191 ± 3,057 and was homogeneous with respect to molecular weight. This product was characterized in regard to physical constants and chemical composition. In general, the molecular properties of human prothrombin complex are very similar to the comparable bovine product. In some preparations a reversible proteolytic enzyme inhibitor (p-aminophenylarsonic acid) was employed in the ultrafiltration step of the purification scheme to inhibit protein degradation.

IMPROVEMENT OF THE SEPTIC LUNG LESION BY TREATMENT WITH A PROTEOLYTIC ENZYME INHIBITOR

1982 ◽  
Vol 57 (3) ◽  
pp. A132-A132
Author(s):  
D. L. Traber ◽  
L. Sziebert ◽  
T. Adams ◽  
N. Henriksen ◽  
L. D. Traber
Keyword(s):  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Lung Lesion ◽  
Proteolytic Enzyme Inhibitor

THE USE OF PROTEOLYTIC ENZYME INHIBITOR (TRASYLOL) IN RADIORECEPTOR ASSAY OF TSH - ITS APPLICATION TO MEASURING THYROID STIMULATING IMMUNOGLOBULINS (TSI) IN THYROID DISEASES

1978 ◽  
Vol 88 (1) ◽  
pp. 65-74 ◽  
Author(s):  
Michizo Kishihara ◽  
Yoshinobu Nakao ◽  
Yasuto Baba ◽  
Shozo Ohgo ◽  
Shigeru Matsukura ◽  
...  
Keyword(s):  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Great Majority ◽  
Thyroid Stimulating Hormone ◽  
Radioreceptor Assay ◽  
Human Thyroid ◽  
Graves Disease ◽  
Stable Method ◽  
Thyroid Stimulating Immunoglobulins ◽  
Proteolytic Enzyme Inhibitor

ABSTRACT In order to estimate thyroid stimulating immunoglobulins (TSI) in serum, a stable, reproducible and sensitive radioreceptor assay (RRA) capable of detecting 100 μU of thyroid stimulating hormone (TSH) has been developed using a proteolytic enzyme inhibitor (Trasylol), partially purified human TSH and particulate fractions of human thyroid homogenate. The binding of 125I-labelled TSH to the crude thyroid membranes was significantly increased from 2–3 % to 15–20 % in the presence of Trasylol (2000 KIU per tube). Further investigations suggested that Trasylol might inhibit the aggregation of 125I-labelled TSH during incubation with these membranes. With this assay system, the serum immunoglobulins from a great majority of untreated patients with Graves' disease were shown to inhibit the binding of 125I-labelled TSH to those membranes more markedly than those from control subjects. Therefore, this RRA for TSH was considered to provide a sensitive and stable method for detecting TSI.

Action of a Proteolytic Enzyme Inhibitor on Blood Coagulation in Vitro

1967 ◽  
Vol 18 (01/02) ◽  
pp. 190-197 ◽  
Author(s):  
B Blombäck ◽  
Margareta Blombäck ◽  
P Olsson
Keyword(s):  
Blood Coagulation ◽  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Factor V ◽  
Generation System ◽  
Antifibrinolytic Agents ◽  
Coagulation Activity ◽  
Serum Factors ◽  
Proteolytic Enzyme Inhibitor

SummaryThe action of the kallikrein and trypsin inhibitor, Trasylol, has been studied in a three-stage thrombin generation system. It has been found that Trasylol inhibits one or several of the early reactions of blood coagulation. The inhibition of the early stages is dependent on the concentration of inhibitor, serum factors and factor VIII. The activation of factor V by tiger snake venom is not inhibited by Trasylol. The inhibition is most probably on one or several of the reactions preceding the factor V involved step.Some other known antifibrinolytic agents have also been tested for anti-coagulation activity.

A multicenter trial of the use of the proteolytic enzyme inhibitor aprotinin in colorectal surgery

1989 ◽  
Vol 32 (6) ◽  
pp. 505-508 ◽  
Author(s):  
William G. Sheridan ◽  
Ahmed A. Shandall ◽  
John Alexander-Williams ◽  
Michael R. B. Keighley ◽  
Paul B. Boulos ◽  
...  
Keyword(s):  
Colorectal Surgery ◽  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Multicenter Trial ◽  
Proteolytic Enzyme Inhibitor

Effect of a proteolytic enzyme inhibitor on the cardiopulmonary response to endotoxin

Resuscitation ◽  
1986 ◽  
Vol 14 (1-2) ◽  
pp. 91-104 ◽  
Author(s):  
D.L. Traber ◽  
T. Adams ◽  
L. Sziebert ◽  
L.D. Traber
Keyword(s):  
Proteolytic Enzyme ◽  
Enzyme Inhibitor ◽  
Proteolytic Enzyme Inhibitor

THE DISTRIBUTION OF TRYPSIN AND CHYMOTRYPSIN INHIBITORS IN POTATO TUBERS

1989 ◽  
Vol 69 (2) ◽  
pp. 513-515
Author(s):  
J. W. G. NICHOLSON ◽  
J. G. ALLEN
Keyword(s):  
Key Words ◽  
Proteolytic Enzyme ◽  
Enzyme Inhibitors ◽  
Enzyme Inhibitor ◽  
Potato Cultivars ◽  
Potato Tubers ◽  
Potato Peel ◽  
Proteolytic Enzyme Inhibitor ◽  
Chymotrypsin Inhibitors

Potato peel contains trypsin and chymotrypsin inhibitors at levels similar to those in other parts of the tuber. The levels of these inhibitors differ among potato cultivars (P < 0.01). The proteolytic enzyme inhibitors may have to be deactiviated before peel waste can be efficiently utilized by pigs. Key words: Potato steam peel, trypsin, chymotrypsin, proteolytic enzyme inhibitor

scholarly journals The purification in high yield and characterization of rat hepatic glucokinase

1976 ◽  
Vol 153 (2) ◽  
pp. 363-373 ◽  
Author(s):  
M J Holroyde ◽  
M B Allen ◽  
A C Storer ◽  
A S Warsy ◽  
J M E Chesher ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Affinity Chromatography ◽  
Amino Acid Composition ◽  
Specific Activity ◽  
Sedimentation Equilibrium ◽  
High Yield ◽  
Equilibrium Studies ◽  
Enzyme Specific Activity

A new improved procedure for the purification of rat hepatic glucokinase (ATP-d-glucose 6-phosphotransferase, EC 2.7.1.2) is given. A key step is affinity chromatography on Sepharose-N-(6-aminohexanoyl)-2-amino-2-deoxy-d-glucopyranose. A homogeneous enzyme, specific activity 150 units/mg of protein, is obtained in about 40% yield. The molecular weight of the pure enzyme was determined by several procedures. In particular, sedimentation-equilibrium studies under a variety of conditions indicate a molecular weight of 48000 and no evidence for dimerization; reports in the literature of other values are discussed in the light of this evidence on the pure enzyme. The amino acid composition suggests that hepatic glucokinase is closely related to rat brain hexokinase and also the wheat “light” hexokinases.

scholarly journals Platelet-derived growth factor. Isolation by a large-scale procedure and analysis of subunit composition

1981 ◽  
Vol 193 (3) ◽  
pp. 907-913 ◽  
Author(s):  
C H Heldin ◽  
B Westermark ◽  
A Wasteson
Keyword(s):  
Molecular Weight ◽  
Growth Factor ◽  
Gel Electrophoresis ◽  
Large Scale ◽  
Specific Activity ◽  
Sodium Dodecyl ◽  
High Specific Activity ◽  
Equilibrium Analysis ◽  
Platelet Derived Growth Factor ◽  
Sedimentation Equilibrium

Platelet-derived growth factor was purified from fresh platelets by a large-scale procedure not involving the use of SDS (sodium dodecyl sulphate). The product, 0.5 mg of platelet-derived growth factor, obtained from about 3 × 10(13) platelets migrated as a single component in analytical gel electrophoresis in the presence of SDS and showed no inhomogeneity on sedimentation-equilibrium analysis in the ultracentrifuge. It had a high specific activity, 2 ng of platelet-derived growth factor/ml (70pM) being equivalent to 1% (v/v) human serum in an assay for multiplication-stimulating activity. Amino acid analysis revealed that platelet-derived growth factor contains all the common amino acids, except tryptophan, but no hexosamine. The molecular weight of platelet-derived growth factor, as determined by sedimentation-equilibrium analysis, was about 33 000. A similar value was obtained by gel electrophoresis in SDS under non-reducing conditions. In the presence of reducing agents the factor molecule was converted into two distinct components of lower molecular weight (17 000 and 14 000 respectively), as demonstrated by protein staining. The molecular model implicated by these findings is that platelet-derived growth factor consists of two different polypeptides chains, linked by disulphide bridges.

Sign in / Sign up

Export Citation Format

Share Document