Some Characteristics of the Clottable Protein of Limulus Polyphemus Blood Cells

1970 ◽  
Vol 23 (01) ◽  
pp. 170-181 ◽  
Author(s):  
N. O Solum
Keyword(s):  
Disc Electrophoresis ◽  
Cell Protein ◽  
Total Amino Acid ◽  
Molecular Characteristics ◽  
Buffer System ◽  
Starch Gel Electrophoresis ◽  
Cell Extracts ◽  
Minimal Molecular Weight ◽  
Acid Buffer System ◽  
Starch Gel

Summary1. The endotoxin-clottable protein of Limulus blood cell extracts has been studied. The concept of the clottable protein as a true cell protein was confirmed.2. 35–55% of total extractable protein was removed from the extracts by clotting. Polyacrylamide disc electrophoresis of the extracts showed 6 to 9 protein bands one of which was reduced in intensity by the clotting. Dimethylf ormamide could be used for fractionated precipitation of the proteins.3. The gel protein was easily soluble in HCl or NaOH and reprecipitated by neutralization. It was also soluble in 0.05 M formate/6.7 M urea pH 4.3 but not in neutral solutions of urea.4. Light absorption spectra of the gel protein in 0.188 N NaOH showed maxima at 283 mμ and 290 mμ, whereas one maximum, at 276 mμ, was observed in 0.189 N HCl. E1 % 1 cm in 0.188 N NaOH was 10.4 and 11.1 at 283 mμ and 290 mμ, respectively, and 9.0 at 276 mμ in 0.189 N HCl.5. Data on the total amino acid composition of the gel protein are given. A mean minimal molecular weight of about 20,000 is calculated from these.6. In starch gel electrophoresis with a discontinuous acid buffer system the gel protein separated into two main zones. Possible relationships between these are discussed in terms of clotting mechanism.7. The data show that Limulus clottable protein differs markedly in its molecular characteristics from those of mammalian fibrinogens.

Bovine Platelet Proteins III. Some Properties of Platelet Fibrinogen

1969 ◽  
Vol 21 (03) ◽  
pp. 428-440 ◽  
Author(s):  
N. O Solum ◽  
S Łopaciuk
Keyword(s):  
Intrinsic Viscosity ◽  
Rabbit Antiserum ◽  
Disc Electrophoresis ◽  
Carbohydrate Content ◽  
Plasma Fibrinogen ◽  
Total Amino Acid ◽  
Electrophoretic Mobilities ◽  
Bovine Plasma ◽  
Starch Gel ◽  
The Difference

Summary1. Some properties of purified bovine platelet fibrinogen have been described and the data compared to those obtained by parallel analysis of purified bovine plasma fibrinogen.2. A close similarity was found between platelet and plasma fibrinogen as to sedimentation coefficients, electrophoretic mobilities in starch gel and polyacrylamide disc electrophoresis, light absorption spectra in the range 240 mμ to 330 mμ, ability to form immunoprecipitate with a rabbit antiserum against bovine plasma fibrinogen, total amino acid composition and in N-terminal amino acids.Differences between the fibrinogens were found as to intrinsic viscosity, carbohydrate content and behaviour upon clotting by thrombin. Intrinsic viscosity in 0.3 M NaCl at 25° was 0.48 dl/g for platelet fibrinogen as compared to 0.26 dl/g for plasma fibrinogen. The carbohydrate content of platelet fibrinogen was 0.56 ± 0.10% 1.56±0.10% and 1.37±0.09% for sialic acid (calculated as N-glycolyl neuraminic acid), hexose (galactose/mannose 1:2) and hexosamine (glucosamine), respectively. These values were 6, 54 and 26% higher than those found for plasma fibrinogen. The difference in clotting behaviour indicated a slower polymerization rate of the fibrin monomers formed from platelet fibrinogen than of those formed from plasma fibrinogen.

Bovine Platelet Proteins II. Purification of Platelet Fibrinogen

1969 ◽  
Vol 21 (03) ◽  
pp. 419-427 ◽  
Author(s):  
N. O Solum ◽  
S Łopaciuk
Keyword(s):  
Analytical Ultracentrifugation ◽  
Insoluble Fraction ◽  
Disc Electrophoresis ◽  
Plasma Fibrinogen ◽  
Starch Gel Electrophoresis ◽  
Freezing And Thawing ◽  
Insoluble Material ◽  
High Resolution Power ◽  
Starch Gel ◽  
Platelet Proteins

Summary1. Platelet fibrinogen has been purified from washed bovine platelets. The procedure was based on the methods for purification of plasma fibrinogen by fractionated precipitations and extractions with ethanol and glycine below 0°, and precipitation of proteins by dimethylformamide at 0°.2. The platelet extract obtained by freezing and thawing of the cells, freed from insoluble material by centrifugation at 23,000 x g for 30 min, contained 0.22 ±0.003mg fibrinogen per 109 platelets. Total protein of this fraction was 0.77 ±0.08 mg per 109 platelets whereas that of the insoluble fraction was 0.79 ±0.09 mg per 109 platelets.3. The most purified platelet fibrinogen fraction contained 91-98% of the protein in a thrombin-clottable state. The yield was approx. 20%. It showed homogeneity in analytical ultracentrifugation, in immunoelectrophoresis using an antiserum produced by immunization of rabbits against platelet extract, and in starch gel electrophoresis using a discontinuous system of Tris HCl and borate buffers offering a high resolution power towards the platelet proteins. Polyacrylamide disc electrophoresis revealed two to three faint lines behind the main fibrinogen line. At least one such line was also observed with purified plasma fibrinogen.

A New Method for Starch Gel Electrophoresis of Human Hemoglobins, with Special Reference to the Determination of Hemoglobin A2

1958 ◽  
Vol 4 (6) ◽  
pp. 484-495 ◽  
Author(s):  
C A J Goldberg
Keyword(s):  
Special Reference ◽  
Gel Electrophoresis ◽  
New Method ◽  
Resolving Power ◽  
Buffer System ◽  
Starch Gel Electrophoresis ◽  
Healthy Individuals ◽  
Starch Gel ◽  
Hemoglobin A2

Abstract A method for starch gel electrophoresis of hemoglobins is presented in which a modified Lintner starch is used for the preparation of the gel. A discontinuous buffer system of tris-EDTA-borate/barbital is used as the electrolyte medium because of its superior resolving power. Hemoglobin A2 values, obtained with this method, of healthy individuals, patients with thalassemia, and those with various anemias of nonthalassemic origin are presented.

Inter-Species Relationships Within the Genus Oncorhynchus Based on Biochemical Systematics

1966 ◽  
Vol 23 (1) ◽  
pp. 101-107 ◽  
Author(s):  
H. Tsuyuki ◽  
E. Roberts
Keyword(s):  
Phylogenetic Relationships ◽  
Gel Electrophoresis ◽  
Plasma Proteins ◽  
Disc Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Species Relationships ◽  
Oncorhynchus Masou ◽  
Starch Gel ◽  
Specific Muscle ◽  
Species Specific

The species specific muscle myogens of Salmo gairdnerii, Oncorhynchus masou, O. masou ishikawae, O. kisutch, O. tshawytscha, O. keta, O. nerka, and O. gorbuscha are compared by starch gel electrophoresis. Plasma proteins of these same species are also examined by polyacrylamide disc electrophoresis. The range of usefulness of muscle myogens in species identification, and equally significantly, their value in establishing phylogenetic relationships of closely related groups, as the genus Oncorhynchus, are discussed. The myogen patterns of O. keta and O. gorbuscha from the Asiatic and North American coasts were found to be identical, further supporting the concept of absolute species specificity of these patterns.

Comparative Zone Electropherograms of Muscle Myogens and Blood Proteins of Adult and Ammocoete Lamprey

1967 ◽  
Vol 24 (6) ◽  
pp. 1269-1273 ◽  
Author(s):  
J. F. Uthe ◽  
H. Tsuyuki
Keyword(s):  
Great Lakes ◽  
Gel Electrophoresis ◽  
Plasma Proteins ◽  
Disc Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Blood Proteins ◽  
Adult Form ◽  
Starch Gel

Polyacrylamide disc electrophoresis of plasma proteins and starch-gel electrophoresis of hemoglobins and muscle myogens of adult and ammocoete forms of three species of Great Lakes lamprey were carried out. Blood proteins were shown to undergo marked changes upon transformation of the ammocoete into the adult form. Muscle myogen did not undergo any change during transformation. The muscle myogen electropherograms of Ichthyomyzon unicuspis and Lampetra lamottei were found to be the same.

scholarly journals Inheritable Differences in the Electrophoretic Pattern of Hemoglobin Revealed in a Local Random-Bred Colony of Albino Rats

Blood ◽  
1970 ◽  
Vol 35 (4) ◽  
pp. 447-450 ◽  
Author(s):  
JOVO V. MARTINOVIC ◽  
DOBRIVOJE V. MARINKOVIC ◽  
DUSAN T. KANAZIR ◽  
PETER N. MARTINOVITCH
Keyword(s):  
Gel Electrophoresis ◽  
Electrophoretic Pattern ◽  
Albino Rats ◽  
Buffer System ◽  
Starch Gel Electrophoresis ◽  
Parental Type ◽  
Starch Gel ◽  
F2 Generation

Abstract In a local colony of random-bred Albino rats, three different patterns of hemoglobin, arbitrarily denoted as patterns I, II and III, were detected by means of starch-gel electrophoresis in a discontinuous buffer system. Mating experiments showed that rats bearing pattern I and pattern III hemoglobin bred true. Crosses of pattern I with pattern III animals yielded only pattern II animals, and when the latter were mated inter se, the resultant F2 generation showed approximately a ratio of 1:2:1 for patterns I, II and III, respectively. When F1 animals from pattern I with pattern III crosses were mated back to animals of either parental type, the resultant ratio was found to be one pattern II: one pattern I or pattern III.

scholarly journals HETEROGENEITY OF THE INHERITED GROUP-SPECIFIC COMPONENT OF HUMAN SERUM

1964 ◽  
Vol 120 (1) ◽  
pp. 83-91 ◽  
Author(s):  
Alexander G. Bearn ◽  
F. David Kitchin ◽  
Barbara H. Bowman
Keyword(s):  
Human Serum ◽  
Gel Electrophoresis ◽  
Polypeptide Chain ◽  
Buffer System ◽  
Starch Gel Electrophoresis ◽  
Specific Component ◽  
Agar Gel ◽  
Main Band ◽  
Starch Gel ◽  
Normal Human

Heterogeneity of the group-specific (Gc) components in normal human serum has been demonstrated by the use of a lithium borate buffer system in conventional vertical starch gel electrophoresis and by prolonged immunoelectrophoresis in agar gel. In both Gc 1-1 and Gc 2-2 phenotypes a protein component migrates ahead of the main band. Immunological evidence indicates that the faster migrating band contains Gc specificity. The possibility that the two electrophoretically distinct Gc components share a common polypeptide chain is discussed.

scholarly journals A Fetal Hemoglobin with Abnormal γ-Polypeptide Chains: Hemoglobin Warren

Blood ◽  
1965 ◽  
Vol 26 (5) ◽  
pp. 668-676 ◽  
Author(s):  
T. H. J. HUISMAN ◽  
A. M. DOZY ◽  
B. E. HORTON ◽  
J. B. WILSON
Keyword(s):  
Fetal Hemoglobin ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Total Amino Acid ◽  
Starch Gel Electrophoresis ◽  
Spectral Absorption ◽  
Abnormal Hemoglobin ◽  
Starch Gel ◽  
Polypeptide Chains

Abstract The discovery of a variant of fetal hemoglobin in the cord blood of a Negro newborn is reported. The abnormal hemoglobin, designated as Hb-Warren, was studied by different analytic procedures, such as starch gel electrophoresis, anion exchange chromatography, ultraviolet spectral absorption, immunologic reactivity with various antisera, hybridization with canine hemoglobin and the determination of the total amino acid composition. It was concluded that Hb-Warren was composed of normal α-polypeptide chains and abnormal γ-polypeptide chains.

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