Functional Relationship of Polymerization Sites of Vertebrate Fibrinogens

1979 ◽  
Author(s):  
C Cierniewski ◽  
T Krajewski ◽  
E Janiak
Keyword(s):  
Gel Electrophoresis ◽  
Functional Relationship ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Functional Similarity ◽  
Fractionation Method ◽  
Relationship Of ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule ◽  
Cold Ethanol Fractionation

Various studies on the interaction of immobilized mammalian fibrinogen and fibrin monomers with some fibrinogen derivatives demonstrated the presence of two sets of polymerization sites in the mammalian fibrinogen molecule. We obtained the same results while investigating the fibrinogen molecules of other classes of vertebrates /Pisces. Amphibia. Aves/. Despite significant differences among their subunit structures, all of them contain polymerization sites homologous to mammalian counterparts. Moreover, due to great functional similarity, fibrinogen or fibrin monomers of the analyzed species of Pisces. Amphibia. Aves and Mammalia interacted in a specific way with immobilized pig fibrin monomers or fibrinogen, respectively. Using these pig affinity adsorbents, fibrinogen and fibrin monomers of different vertebrates were isolated directly from plasma and analyzed by SDS polyacrylamide gel electrophoresis. Polypeptide compositions of eluted proteins were identical to those obtained for corresponding fibrinogen preparations isolated by cold-ethanol fractionation method. It appears to indicate that the nature of polymerization sites in vertebrate fibrinogens is alike.

Functional Relationship of Polymerization Sites of Vertebrate Fibrinogens

1979 ◽  
Author(s):  
C.S. Ciernlewski ◽  
T. Krajewski ◽  
E. Janiak
Keyword(s):  
Gel Electrophoresis ◽  
Functional Relationship ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Functional Similarity ◽  
Fractionation Method ◽  
Relationship Of ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule ◽  
Cold Ethanol Fractionation

Various studies on the interaction of immobilized mammalian fibrinogen and fibrin monomers with some fibrinogen derivatives demonstrated the presence of two sets of polymerisation sites In the mammalian fibrinogen molecule. We obtained the same results while investigating the fibrinogen molecules of other classes of vertebrates /Pisces, Amphibia, Aves/. Despite significant differences among their subunit structures, all of them contain polymerization sites homologous to mammalian counterparts. Moreover, due to great functional similarity, fibrinogen or fibrin monomers of the analyzed species of Pisces, Amphibia, Ayes and Mammalia interacted in a specific way with Immobilized pig fibrin monomers or fibrinogen, respectively. Using these pig affinity adsorbents, fibrinogen and fibrin monomers of different vertebrates were isolated directly from plasma and analyzed by SDS Polyacrylamide gel electrophoresis. Polypeptide compositions of eluted proteins were identical to those obtained for corresponding fibrinogen preparations isolated by cold-ethanol fractionation method. It appears to indicate that the nature of polymerization sites in vertebrate fibrinogens is alike.

A New Fibrinogen Variant With Abnormal Gamma Chain : Fibrinogen Haifa

1981 ◽  
Author(s):  
J Soria ◽  
C Soria ◽  
G Palareti ◽  
M Tavori ◽  
M Samama ◽  
...  
Keyword(s):  
Protective Effect ◽  
Gel Electrophoresis ◽  
Arterial Thrombosis ◽  
Degradation Products ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Gamma Chain ◽  
Peripheral Arterial ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule

A congenital abnormal fibrinogen was detected in a 30 year old woman presenting several episodes of peripheral arterial thrombosis.The abnormality in fibrin formation is located in the fibrin monomers aggregation, since the fibrinopeptides release and the fibrin stabilization were normal.The SDS polyacrylamide gel electrophoresis of plasmic degradation products, obtained either in the presence or in the absence of calcium, revealed an absence of protective effect of calcium for plasmin degradation of Fibrinogen Haifa. Since, Ca++ protects against further plasmin degradation of the gamma chain, we suggest that the anomaly is located in this part of the fibrinogen molecule.Because the anomaly was discovered at Haifa, Israel, we suggest to call this abnormal fibrinogen : “Fibrinogen Haifa”.

An Abnormal Fibrinogen (Copenhagen) Associated with Severe Thromboembolic Disease, but with Normal Adsorption of Plasminogen

1979 ◽  
Author(s):  
Sandbjerg M. Hansen ◽  
I. Clemmensen
Keyword(s):  
Venous Thrombosis ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Thromboembolic Disease ◽  
Polyacrylamide Gel ◽  
Fibrinopeptide A ◽  
Related Material ◽  
Thrombotic Disease ◽  
Fibrin Formation ◽  
Fibrin Monomers

An autosomally inherited qualitative fibrinogen (F) defect is presented. The abnormal F was detected in the plasma of a 54 year old woman with severe arterial thrombotic disease. A decreased rate of fibrin formation of plasma, or purified F by thrombin or ancrod (Arvin (R ) ) was demonstrated. The plasma F concentration was normal, when estimated by clottability or immunologic technique. No F related material was present in the serum. The purified abnormal F was indistinguishable from normal F by Immunoelectrophoresis or SDS Polyacrylamide gel electrophoresis. The major defects detected were an abnormality of polymerization of fibrin monomers and a decreased rate of release of fibrinopeptide A. To evaluate a possible abnormality of the binding of plasminogen (P) to the abnormal fibrin, we examined the adsorption of partially degraded P (Lys-P), which has a higher affinity for fibrin than Glu-P. The adsorption was normal, but the study might be useful in the evaluation of dysfibrinogenemia associated with venous thrombosis.

scholarly journals Protein extraction method of Metroxylon sagu leaf for high resolution two dimensional gel electrophoresis and comparative proteomics

2019 ◽  
Author(s):  
Mehvish Nisar ◽  
Hasnain Hussain
Keyword(s):  
Gel Electrophoresis ◽  
Extraction Method ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Fractionation Method ◽  
Metroxylon Sagu ◽  
Time Required

Abstract Objective This study aimed to determine the best protein extraction method of Metroxylon sagu for the two-dimensional gel electrophoresis and the comparative analysis. Results To perform good proteome research, the most critical step is to establish a method that gives the best quality of extracted total proteins. To develop an optimized protein extraction protocol for two-dimensional polyacrylamide gel electrophoresis (2-DE) analysis of Metroxylon sagu, five protein extraction protocols were compared; polyethene glycol (PEG) fractionation method, SDS/phenol method, TCA/acetone method, combination SDS/phenol and TCA/acetone and imidazole method. The PEG fractionation method was found to give the most reproducible gels with the highest number of spots and highest protein concentration followed by SDS/Phenol method. The lowest number of spots were observed in Imidazole method. The PEG fractionation method provides improved resolution and reproducibility of two-dimensional polyacrylamide gel electrophoresis (2-DE) and reduces the time required to analyze samples. Partitioning rubisco by polyethene glycol (PEG) fractionation provides clearer detection of low-abundance protein. Hence the result from this study propose PEG fractionation as the effective protein extraction method for 2-DE proteomic studies of Metroxylon sagu.

Soluble Fibrin Complexes: Separation as a Function of pH and Characterization

1977 ◽  
Vol 37 (01) ◽  
pp. 144-153 ◽  
Author(s):  
Y Benabid ◽  
E Concord ◽  
M Suscillon
Keyword(s):  
Gel Electrophoresis ◽  
Polypeptide Chain ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Chromatography ◽  
Elution Buffer ◽  
Chain Composition ◽  
Two Peaks ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule ◽  
High Polymers

Summary1. The influence of the pH on the separation of high molecular weight derivatives obtained by a limited action of thrombin on fibrinogen was studied by agarose gel chromatography. When the pH of the elution buffer was 8.5, non crosslinked associations were easily separated in two peaks eluted prior to the fibrinogen peak: one contained a dimer, the other several high polymers. At pH 6.5, only the fibrinogen peak appeared: the fibrinogen molecule proteolysed by thrombin formed no stable associations at this pH and was eluted with the intact fibrinogen molecule. In the presence of factor XIII and Ca++, numerous associations were obtained which are independant of the pH.2. The polypeptide chain composition of the different species separated at pH 8.5 was studied by SDS-polyacrylamide gel electrophoresis. This technic showed Aα, Bβ and γ chains in the fibrinogen peak, whereas in the chromatographic fractions containing the dimer four bands corresponding to Aα, α, Bβ and γ chains were found. In the peak containing the high polymers, only the presence of α, Bβ and γ chains was demonstrated.3. These experimental results concerning the effect of pH on the formation of soluble complexes showed that the presence of fibrin monomers in fibrinogen solution was not sufficient to promote any associations. The formation of such derivatives is strongly dependent on the pH of the solution. This obviously can be explained by an influence of the pH either on the ionization of polymerisation sites and the intermolecular bonds between the complex units or on the unmasking of the polymerisation sites by a hypothetical pH induced conformational change of the fibrinogen molecule.

Two-dimensional SDS–polyacrylamide gel electrophoresis of heat-modifiable outer-membrane proteins

1976 ◽  
Vol 22 (1) ◽  
pp. 83-91 ◽  
Author(s):  
R. R. B. Russell
Keyword(s):  
Gel Electrophoresis ◽  
Outer Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Two Dimensional ◽  
Two Temperatures ◽  
Cell Envelopes ◽  
Neisseria Sicca ◽  
Relationship Of ◽  
The Relationship

An examination has been made of the effect which temperature of solubilization has upon the subsequent migration in SDS–polyacrylamide gel electrophoresis of proteins from the cell envelopes of Escherichia coli K12 and Neisseria sicca ATCC 9913. Conventional electrophoresis in tubes revealed substantial differences in the staining patterns of gels, depending upon whether the envelope samples were solubilized at 37 °C or 100 °C; in the case of N. sicca at least 6 of 13 discernible bands displayed heat-modifiable behavior. The relationship of the bands produced by each of the two temperatures was investigated by a two-dimensional electrophoresis procedure, in which a sample was solubilized at 37 °C and run in a usual cylindrical gel; the entire gel was then resolubilized at 100 °C, and laid along an acrylamide slab for electrophoresis in the second dimension.It was found that "free endotoxin" of both organisms examined contained the same major proteins as the total envelope fraction, and that these free endotoxin proteins showed the same heat-modifiable properties as when present in total envelopes.

scholarly journals Protein extraction method of Metroxylon sagu leaf for high resolution two dimensional gel electrophoresis and comparative proteomics

2019 ◽  
Author(s):  
Mehvish Nisar ◽  
Hasnain Hussain
Keyword(s):  
Gel Electrophoresis ◽  
Extraction Method ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Fractionation Method ◽  
Metroxylon Sagu ◽  
Time Required

Abstract Objective This study aimed to determine the best protein extraction method of Metroxylon sagu for the two-dimensional gel electrophoresis and the comparative analysis. Results To perform good proteome research, the most critical step is to establish a method that gives the best quality of extracted total proteins. To develop an optimized protein extraction protocol for two-dimensional polyacrylamide gel electrophoresis (2-DE) analysis of Metroxylon sagu, five protein extraction protocols were compared; polyethene glycol (PEG) fractionation method, SDS/phenol method, TCA/acetone method, combination SDS/phenol and TCA/acetone and imidazole method. The PEG fractionation method was found to give the most reproducible gels with the highest number of spots and highest protein concentration followed by SDS/Phenol method. The lowest number of spots were observed in Imidazole method. The PEG fractionation method provides improved resolution and reproducibility of two-dimensional polyacrylamide gel electrophoresis (2-DE) and reduces the time required to analyze samples. Partitioning rubisco by polyethene glycol (PEG) fractionation provides clearer detection of low-abundance protein. Hence the result from this study propose PEG fractionation as the effective protein extraction method for 2-DE proteomic studies of Metroxylon sagu.

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