scholarly journals Congenital Coagulation Factor IX, Protein C and Antithrombin III Structural Aspects from Three Dimensional Model

1997 ◽  
Vol 8 (2) ◽  
pp. 84-102
Author(s):  
Toshiyuki MIYATA ◽  
Hiroaki TSUKAMOTO ◽  
Koji SUZUKI ◽  
Hideaki UMEYAMA
Keyword(s):  
Protein C ◽  
Antithrombin Iii ◽  
Three Dimensional ◽  
Coagulation Factor ◽  
Factor Ix ◽  
Dimensional Model ◽  
Three Dimensional Model ◽  
Coagulation Factor Ix ◽  
Structural Aspects

EXPRESSION IN ONTOGENESIS OF HUMAN BLOOD COAGULATION FACTORS

1987 ◽  
Author(s):  
H J Hassan ◽  
A Leonardi ◽  
C Chelucci ◽  
R Guerriero ◽  
P M Mannucci ◽  
...  
Keyword(s):  
Blood Coagulation ◽  
Protein C ◽  
Antithrombin Iii ◽  
Liver Homogenate ◽  
Coagulation Factor ◽  
Coagulation Factors ◽  
Factor Ix ◽  
Adult Liver ◽  
Blood Coagulation Factors ◽  
Guanidine Isothiocyanate

We have analyzed the expression of several blood coagulation factors (IX, VIII, X, fibrinogen chains) and inhibitors (antithrombin III, protein C) in human embryonic and fetal livers, obtained from legal abortions at 6-11 week post-conception. The age was established by morphologic staging and particularly crown-rump lenght measurement.Total cellular RNA was isolated from partially purified hepatocytes or total liver homogenate using the guanidine isothiocyanate method. Poly(A)+ RNA was selected by oligodT cellulose chromatography. The size and the number of the embryonic and fetal transcripts are equivalent to those observed in adult liver, as evaluated by Northern blot analysis of total or poly(A)+ RNA hybridized to human cDNA probes.The level of coagulation factor transcripts in embryonic and fetal liver was evaluated by dot hybridization of total RNA (0.5-10 ug), as compared to RNA extracted from normal adult liver biopsies. The expression of blood coagulation factors in embryos is generally reduced for all factors, but at a different degree. In 5-11 wk liver, the level of factor IX is 5-10% of that observed in adults, while fibrinogen, protein C, antithrombin III RNA level rises from 25 to 50% and factor X is expressed at a level comparable to that observed in adult liver.We conclude that during these stages of development blood coagulation factors are expressed according to three different time, curves, possibly due to the effect of different types of regulatory mechanisms.

Construction, Expression, and Properties of a Recombinant Chimeric Human Protein C with Replacement of Its Growth Factor-like Domains by Those of Human Coagulation Factor IX

Biochemistry ◽  
1994 ◽  
Vol 33 (3) ◽  
pp. 823-831 ◽  
Author(s):  
Shiqin Yu ◽  
Li Zhang ◽  
Ashish Jhingan ◽  
William T. Christiansen ◽  
Francis J. Castellino
Keyword(s):  
Growth Factor ◽  
Protein C ◽  
Coagulation Factor ◽  
Factor Ix ◽  
Human Protein ◽  
Coagulation Factor Ix

Immunopurification of Human Coagulation Factor IX Using Monoclonal Antibodies

1986 ◽  
Vol 56 (01) ◽  
pp. 086-089 ◽  
Author(s):  
Hagop Bessos ◽  
Christopher V Prowse
Keyword(s):  
Monoclonal Antibodies ◽  
Protein C ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Coagulation Factor ◽  
Factor Ix ◽  
Coagulation Factor Ix ◽  
Immunoaffinity Purification ◽  
Sds Page ◽  
Sepharose 4B

SummaryThis study examines the suitability of four recently characterised monoclonal antibodies (MAbs) for the immunoaffinity purification of human coagulation factor IX (FIX) from plasma concentrates. Initial studies using 125I-FIX indicated that appreciable amounts of bound FIX could be eluted from immobilised MAbs with 0.2 M glycine, 50% ethanediol pH 10 (buffer N). Further studies with FIX concentrates showed that buffer N eluted FIX without compromising the activity of the zymogen. Although FIX was eluted from all four MAbs with this buffer, the best yield (82%) was obtained with MAb ESN-3. ESN-3 bound 40 to 60 iu FIX per mg MAb when immobilised on Sepharose 4B. After washing, column elution with buffer N yielded FIX at 100-200 iu/mg. The purity of the product was confirmed by sodium-dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting. The product contained no detectable mouse IgG (<3%) and <1% FII, X, or protein C.

Three-dimensional Model of Coagulation Factor Va Bound to Activated Protein C

2000 ◽  
Vol 84 (11) ◽  
pp. 849-857 ◽  
Author(s):  
Jean-Luc Pellequer ◽  
Andrew Gale ◽  
Elizabeth Getzoff ◽  
John Griffin
Keyword(s):  
Protein C ◽  
Molecular Model ◽  
Three Dimensional ◽  
Activated Protein C ◽  
Coagulation Factor ◽  
Factor V ◽  
C2 Domain ◽  
Phospholipid Membrane ◽  
Three Dimensional Model ◽  
Factor Va

SummaryA complete molecular model of blood coagulation factor Va (FVa) bound to anticoagulant activated protein C (APC) and to a phospholipid membrane was constructed. The three homologous A domains and the two homologous C domains of FVA were modeled based on the X-ray crystallographic structures of ceruloplasmin and C2 domain of factor V, respectively. The final arrangement of the five domains in the complete FVa model bound to a membrane incorporated extensive published experimental data. FVa binds the phospholipid membrane through its C2 domain while the A-domain trimer is located from 40 through 100 Å above the membrane plane. From our model we infer a probable role for metal ions at the interface between FVa light and heavy chains, provide an explanation for the slower APC cleavage at Arg306 relative to Arg506, and predict specific interactions between positively and negatively charged exosites in APC and FVa, respectively.

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