Sialic Acid Dependent Polypeptide Chain Heterogeneity of Human Fibrinogen Demonstrated by Two-Dimensional Electrophoresis

A dysfibrinogenemia (fibrinogen Sevilla) was detected in a 64-yr-old woman with no previous history of hemorrhagic diathesis or thrombosis. Thrombin and reptilase times were prolonged. The aggregation of fibrin monomers showed a prolonged latency time with a defective slope although fibrinopeptide release and clot stabilization were found to be normal. Plasmin proteolysis was abnormal with a much slower plasmic degradation in patient's purified fibrinogen. By chromatofocussing the patient's fibrinogen showed an abnormality in pattern elution with a second peak eluting at a pH slightly more basic than the normal one @H 5.5). Likewise, the isoelectrofocussing of purified non-reduced patient's fibrinogen in agarose gel showed an abnormal distribution in its focussed bands, especially in a group which focussed in a p&interval between 5.20-5.85. By two-dimensional electrophoresis we did not find any abnormality in the fibrinogen-reduced chains. These results could indicate that the abnormal monomer aggregation, as well as the defective plasmin lysis, could be due to conformational aspects of fibrinogen rather than to structural defects.

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“…10). The pattern was similar to that shown by the reduced normal fibrinogen and to that described by other authors [15,16].…”

Section: Isoelectrofocussing In Agarose Gelsupporting

confidence: 91%

Fibrinogen Sevilla, a congenital dysfibrinogenemia characterized by an abnormal monomer aggregation and a defective plasmin lysis

Fernández

Noguerol

Sosa

et al. 1989

Clinica Chimica Acta

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“…In summary, the improved two-dimensional IEFISDS-PAGE which is described here offers a very sensitive means for examining the heterogeneity of the subunits of fibrinogen and for detecting changes in heterogeneity, which may occur both in vivo and in vitro. A similar technique, recently described by Kuyas et al (9) demonstrates similar charge heterogeneity within the subunit chains of fibrinogen, although the variant y chain is not clearly depicted in that system. In addition, the ranges of apparent isoelectric points for the individual chains are different, possibly due to a different method for measuring the pH.…”

Section: Resultsmentioning

confidence: 83%

“…The observed heterogeneity has been reported to result from variations in size in the a chain, supposedly due to in vivo proteolytic cleavages (9, and to both charge (6) and size differences (7) in the y chain. Because such variations are not easily detectable with ordinary electrophoretic techniques, two-dimensional electrophoresis has been used to examine them (3,8,9). However, the Aa and Bp chains are difficult to resolve with isoelectric focusing due to the overlap in the range of isoelectric points.…”

Section: Introductionmentioning

confidence: 99%

High Resolution Electrophoretic Analysis of Human Fibrinogen and Its Crosslinked Intermediates

Holahan

et al. 1983

Thromb Haemost

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SummaryHeterogeneity in human fibrinogen was examined using an improved two-dimensional isoelectric focusing-SDS polyacrylamide gel electrophoretic procedure. Four different preparations of fibrinogen were compared: single donor fibrinogen prepared from plasma by precipitation with ammonium sulfate or by affinity chromatography on fibrin-monomer Sepharose, fraction I—4 prepared from Cohn fraction I paste, and Kabi grade L. The subunit Aα, Bβ, and γ chains in all preparations had marked charge heterogeneity. The three chains were clearly separated from each other and a range of isoelectric points for each chain could be assigned. Minor variations in the subunit heterogeneity of the different preparations were found. Intermediates in the transition from fibrinogen to crosslinked fibrin were also examined. A striking increase in the heterogeneity of the α chain was observed during crosslinking.

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“…We have therefore concluded that measurement of this fraction of fibrinogen may prove to be of clinical diagnostic significance. The present study is an attempt to characterize and explain the heterogeneity of human fibrinogen ( − ). Electrophoretic separation of human serum proteins does not give the estimation for their classification or stability, but it is an additional index for their partial characterization.…”

Section: Resultsmentioning

confidence: 99%

Structural Changes in Plasma Circulating Fibrinogen after Moderate Beer Consumption as Determined by Electrophoresis and Spectroscopy

Gorinstein

Caspi

Goshev

et al. 2002

J. Agric. Food Chem.

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The effects of short-term moderate beer consumption (MBC) on plasma circulating fibrinogen (PCF) in patients suffering from coronary atherosclerosis were investigated by use of 2-dimensional electrophoresis (2-DE), circular dichroism (CD), and Fourier transform infrared spectroscopy (FT-IR). Forty-eight volunteers after coronary bypass surgery were divided into experimental (EG) and control (CG) groups, each of 24. Patients of the EG group consumed 330 mL of beer/day (about 20 g of alcohol) for 30 consecutive days, and CG volunteers drank mineral water instead of beer. Blood samples were collected before and after the experiment. In 21 out of 24 patients after beer consumption the plasma circulating fibrinogen was compromised: changes in its secondary structure were found. These changes were expressed in relatively low electrophoretic mobility and charge heterogeneity, decrease in alpha-helix and increase in beta-sheet, and in slight shift of amide I and II bands. Our findings indicate that one of the positive benefits of moderate beer consumption is to diminish the production of fibrinogen and its stability, which reduces the potential risk exerted by this protein. Thus, in most of beer-consuming patients some qualitative structural changes in plasma circulating fibrinogen were detected.

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Sialic Acid Dependent Polypeptide Chain Heterogeneity of Human Fibrinogen Demonstrated by Two-Dimensional Electrophoresis

Cited by 14 publications

References 11 publications

Fibrinogen Sevilla, a congenital dysfibrinogenemia characterized by an abnormal monomer aggregation and a defective plasmin lysis

Fibrinogen Sevilla, a congenital dysfibrinogenemia characterized by an abnormal monomer aggregation and a defective plasmin lysis

High Resolution Electrophoretic Analysis of Human Fibrinogen and Its Crosslinked Intermediates

Structural Changes in Plasma Circulating Fibrinogen after Moderate Beer Consumption as Determined by Electrophoresis and Spectroscopy

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