Altered amounts of hemoglobin synthesis in livers of dystrophic hamsters

1977 ◽  
Vol 55 (8) ◽  
pp. 894-900 ◽  
Author(s):  
K. Wrogemann ◽  
N. M. Quilliam ◽  
E. G. Nylen ◽  
M. D. Johnson ◽  
J. F. Clancy Jr.
Keyword(s):  
Gel Electrophoresis ◽  
Protein Fraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Leucine Incorporation ◽  
Major Protein ◽  
Hemoglobin Synthesis ◽  
Histological Sections ◽  
Age Dependent ◽  
Different Levels

In liver supernatants similar amounts of leucine are incorporated into proteins of normal and dystrophic hamsters. However, using a dual labeling technique, a protein fraction is detected which shows different levels of leucine incorporation between normal and dystrophic animals in an age-dependent fashion. The major protein in this fraction comigrates with hemoglobin or its subunits under various conditions of polyacrylamide gel electrophoresis. In animals younger than 3 days there is more synthesis of this protein fraction in dystrophic animals, while at 6 days there is considerably less synthesis of this protein. These changes are paralleled by differences in the number of erythroid foci in histological sections of the livers. There is no evidence for structurally altered hemoglobin in dystrophic hamsters. The significance of this finding and its possible relation to the dystrophy process are not known at present.

scholarly journals Purification, properties and specificity of the restriction endonuclease from Bacillus stearothermophilus

1979 ◽  
Vol 177 (1) ◽  
pp. 49-62 ◽  
Author(s):  
C M Clarke ◽  
B S Hartley
Keyword(s):  
Enzyme Activity ◽  
Restriction Endonuclease ◽  
Dna Cleavage ◽  
Gel Electrophoresis ◽  
Bacillus Stearothermophilus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Activity Restriction

The restriction endonuclease BstI was purified from 70kg of Bacillus stearothermophilus. The final product is at least 97% pure as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; this major protein species co-migrates with the enzyme activity on native polyacrylamide-gel electrophoresis and isoelectric focusing. Pure restriction endonuclease BstI has a subunit mol.wt. of 26,000 and is probably a loosely associated dimer. The enzyme shows maximum activity at pH values between 7 and 9.5, and in the presence of 0.5-2mM-Mg2+. NaCl inhibits the restriction enzyme activity. Restriction endonuclease BstI cleaves DNA in a position identical with that cleaved by endonuclease BamHI (for Bacillus amyloliquefaciens), i.e.: (formula: see text). In the presence of high concentrations of enzyme, DNA cleavage occurs at secondary sites. This side-specificity is enhanced by the addition of glycerol. Preliminary studies indicate that these sites are of the type: (formula: see text).

scholarly journals Polyacrylamide gel electrophoresis of soybean seed proteins

2015 ◽  
Vol 47 (4) ◽  
pp. 371-378 ◽  
Author(s):  
W. Lassocińsk ◽  
J. S. Knypl
Keyword(s):  
Glycine Max ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Soybean Seed ◽  
Seed Proteins ◽  
Soluble Proteins ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Molecular Weights ◽  
Minor Protein

Four major and 14 minor protein bands were detected when total salt soluble proteins of soybean (Glycine max cultivar Warszawska) seed were subjected to polyacrylamide gel electrophoresis under nondissociating conditions, and 16 protein bands were detected under dissociating conditions. Molecular weights of three major protein fractions in PAGE SDS were determined for around 18 500, 36 000 and 80 000 daltons.

Sodium dodecyl sulfate polyacrylamide gel electrophoresis of serum after protein denaturation in the presence or absence of 2-mercaptoethanol.

1984 ◽  
Vol 30 (3) ◽  
pp. 475-479 ◽  
Author(s):  
T Marshall
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Protein Denaturation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Dodecyl Sulfate ◽  
Electrophoretic Techniques

Abstract Human serum proteins were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis after protein denaturation in the presence or absence of 2-mercaptoethanol. Both electrophoretic techniques give characteristic and reproducible banding patterns and achieve a high degree of resolution within the limits of a one-dimensional separation. The major protein bands have been identified, and the merits of the two techniques are compared. Protein denaturation in the absence of 2-mercaptoethanol gives more discrete bands for the purpose of protein identification by maintaining the disulfide-bridge-dependent polypeptide associations characteristic of many serum proteins. However, simultaneous use of both electrophoretic techniques enhances identification by exploiting the response of an individual protein to mercaptoethanol treatment. The clinical potential of this approach for detecting protein disorders is discussed.

scholarly journals Electrophoretic and immunological charactorization of rat neurophysin

1971 ◽  
Vol 122 (5) ◽  
pp. 671-676 ◽  
Author(s):  
A. Norström ◽  
J. Sjöstrand ◽  
B. G. Livett ◽  
L. O. Uttenthal ◽  
D. B. Hope
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Minor Component ◽  
Polyacrylamide Gel ◽  
Male Rats ◽  
Major Protein ◽  
Starch Gel Electrophoresis ◽  
Neural Lobe ◽  
Starch Gel

1. The electrophoretic properties of rat posterior pituitary proteins have been compared on starch gel with those of bovine and porcine neurophysins. 2. [35S]-Cysteine was injected into the supraoptic nucleus of male rats and 16–24h later the distribution of labelled neural-lobe protein in starch and polyacrylamide gels was determined. In both systems a single major protein component was found to contain more than 80% of the total recovered radioactivity. Between 5 and 10% of the radioactivity was found in a minor component in polyacrylamide gel. 3. In agar, microimmuno-diffusion and -electrophoresis of the rat neural-lobe proteins gave a single arc with neurophysin antiserum, and after starch-gel electrophoresis this arc was shown to be due to the major labelled component. 4. The molecular weights of the rat neural-lobe proteins were estimated by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The molecular weight of the major labelled component was found to be 12000. 5. It is concluded that the rat neurophysin consists of one major and possibly one minor component.

scholarly journals Isolation and protein composition of gap junctions from rabbit hearts

1982 ◽  
Vol 205 (1) ◽  
pp. 189-194 ◽  
Author(s):  
C K Manjunath ◽  
G E Goings ◽  
E Page
Keyword(s):  
Gap Junctions ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Composition ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Electron Microscopic ◽  
Gap Junctional

We have modified a method for isolating gap-junctional membrane from mouse hearts [Kensler & Goodenough (1980) J. Cell Biol. 86, 755-764] to isolate gap junctions of comparable purity from rabbit hearts more rapidly, with better yield, and without resort to non-ionic detergents. Purification was monitored by electron microscopy of thin-sectioned membrane pellets and by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Gap junctions were obtained as vesicles whose mean surface area approximated that of junctions in intact myocardial cells. About 10-20% of the vesicles were ferritin-impermeable. Approx. 125 micrograms of membrane protein was obtained per 8 g of rabbit heart. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of purified gap junctions showed five major protein bands of mol.wts. 46 000, 44 000, 33 000, 30 000 and 28 500 that co-purified with the junctions. This protein composition was nearly identical with that published for gap junctions of mouse hearts, and differed markedly from the protein composition of gap junctions from non-excitable cells (lens and liver). The constancy of junctional protein composition between hearts of two different species and its non-identity with that from liver and lens suggest that, although gap-junctional structure in mammalian tissues seems to be remarkably similar by electron-microscopic techniques, junctional-channel protein composition actually varies from tissue to tissue and may be adapted to the permeability requirements of the tissue.

scholarly journals The effect of tunicamycin on the glycosylation of lactating-rabbit mammary glycoproteins

1979 ◽  
Vol 180 (3) ◽  
pp. 481-489 ◽  
Author(s):  
Brian K. Speake ◽  
David A. White
Keyword(s):  
Acid Hydrolysis ◽  
Paper Chromatography ◽  
Gel Electrophoresis ◽  
Protein Fraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Strong Acid ◽  
Polyacrylamide Gel ◽  
Control Value ◽  
Acid Hydrolysate ◽  
Hydrolysis Of

1. Tunicamycin inhibited the incorporation of d-[2-3H]mannose into dolichol-linked oligosaccharide and glycoprotein of lactating-rabbit mammary explants by approximately the same extent (approx. 30% of control value), suggesting that lipid-linked intermediates are involved in the mannosylation of mammary glycoproteins. 2. The incorporation of radioactivity from N-acetyl-d-[1-14C]glucosamine into dolichol-linked oligosaccharide was inhibited by tunicamycin to 32% of the control value, whereas the incorporation of the radiolabel into glycoprotein was only inhibited to 72% of the control value. 3. Considerable redistribution of label from N-acetylglucosamine to N-acetylgalactosamine was found to occur in the explants. In the presence of tunicamycin approx. 76% of the radioactivity incorporated into glycoprotein from N-acetyl-d-[1-14C]glucosamine was present as N-acetylgalactosamine, compared with approx. 61% in the absence of the inhibitor. Thus tunicamycin selectively inhibits the incorporation of N-acetylglucosamine into glycoprotein. 4. Radioactivity from N-acetyl-d-[1-14C]glucosamine was incorporated into a glycoprotein that was identified as casein by the use of a casein-specific antiserum, and also into a group of glycopolypeptides with apparent mol.wts. ranging between 40000 and 80000. N-Acetylgalactosamine was the only radioactive sugar released on strong-acid hydrolysis of the immunoprecipitated casein, whereas N-acetylglucosamine was the major radioactive residue present in the non-casein glycoproteins. Glucosamine and galactosamine were the only radiolabelled sugars detected by paper chromatography of the strong-acid hydrolysate of the protein fraction. 5. Tunicamycin inhibited the incorporation of radioactivity from N-acetyl-d-[1-14C]glucosamine into the glycopolypeptides with mol.wts. between 40000 and 80000 as described by polyacrylamide-gel electrophoresis, but did not affect the incorporation of label into casein. It appears that tunicamycin inhibits the incorporation of mannose and N-acetylglucosamine into a number of mammary glycoproteins by inhibiting the formation of lipid-linked intermediates, but does not inhibit the incorporation of N-acetylgalactosamine into casein.

Protein analysis ofTheileria sergenti/buffeli/orientalispiroplasms by two-dimensional polyacrylamide gel electrophoresis

Parasitology ◽  
1991 ◽  
Vol 102 (3) ◽  
pp. 341-346 ◽  
Author(s):  
C. Sugimoto ◽  
S. Kawazu ◽  
T. Kamio ◽  
K. Fujisaki
Keyword(s):  
Molecular Weight ◽  
Protein Spot ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Analysis ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Two Dimensional ◽  
Characteristic Set

Proteins of the piroplasms ofTheileria sergenti, T. buffeliandT. orientaliswere analysed by two-dimensional polyacrylamide gel electrophoresis. Protein spot patterns ofT. buffeliandT. orientaliswere identical except for a few minor proteins, whereas spot patterns of twoT. sergentistocks were differentiated from those ofT. buffeliandT. orientalisby a characteristic set of proteins including a major protein of molecular weight 33–34 kDa. This result indicates that JapaneseT. sergentican be phenotypically distinguishable from European and AustraliaTheileriaspecies;T. orientalisandT. buffeli.

Differentiation of Rennet from Other Milk-Clotting Enzymes by Polyacrylamide Gel Electrophoresis

1977 ◽  
Vol 60 (6) ◽  
pp. 1372-1374 ◽  
Author(s):  
Manfred J Prager
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
The Other ◽  
Characteristic Pattern ◽  
Aniline Blue ◽  
Major Protein ◽  
Milk Clotting ◽  
Microbial Origin

Abstract Polyacrylamide gel electrophoresis was used to differentiate animal rennet and other milk-clotting enzymes. After electrophoresis, the separated components were visualized by staining with aniline blue-black. Two prominent proteins were found in calf and bovine rennet, while only 1 major protein was observed in pepsin and enzymes of microbial origin. These patterns provided a basis for distinguishing animal rennet and the other enzymes as well as a means of identifying each type of enzyme by the characteristic pattern shown.

scholarly journals Degradation of Bovine Factor VIII by Plasmin and Trypsin

Blood ◽  
1974 ◽  
Vol 43 (5) ◽  
pp. 629-640 ◽  
Author(s):  
Edward P. Kirby ◽  
Neil Martin ◽  
Victor J. Marder
Keyword(s):  
Acetic Acid ◽  
Factor Viii ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Antigenic Determinants ◽  
Major Protein ◽  
Acid System ◽  
Coagulant Activity ◽  
Bovine Factor

Abstract The degradation of bovine Factor VIII by plasmin, trypsin, and thrombin was monitored with polyacrylamide gel electrophoresis in a urea—acetic acid system. The sequences of plasmic and tryptic proteolysis were very similar, proceeding via several intermediate, enzyme-sensitive products to smaller derivatives which were relatively resistant to further degradation. The intermediate degradation products were similar for both enzymes, although prolonged digestion with trypsin produced final products which were smaller than those observed with plasmin. Stages of digestion which reflect the degree of completeness of proteolysis were defined on the basis of the presence or absence of these distinct components in the electrophoretic profiles. Thrombin treatment of Factor VIII caused a loss of coagulant activity but did not yield products small enough to be characterized in this electrophoretic system. During the course of plasmic and tryptic digestion, there was a rapid loss of all coagulant activity without the generation of anticoagulant activity. Agarose chromatography of a 24-hr plasmic digest revealed three major protein peaks, which corresponded to individual electrophoretic bands on polyacrylamide gel electrophoresis in a nondenaturing system at pH 8.8. Upon polyacrylamide gel electrophoresis in the urea—acetic acid system, the components of the first two of these peaks dissociated into two and three separate fragments, respectively. Immunoelectrophoresis demonstrated two major distinct antigenic determinants in these first two elution peaks.

scholarly journals Purification and properties of pyruvate kinase from the hepatopancreas of Carcinus maenas

1976 ◽  
Vol 153 (1) ◽  
pp. 127-134 ◽  
Author(s):  
I G Giles ◽  
P C Poat ◽  
K A Munday
Keyword(s):  
Pyruvate Kinase ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Carcinus Maenas ◽  
Deae Cellulose ◽  
Protein Band ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Major Protein Band

Pyruvatekinase from the hepatopancreas of the common shore crab, Carcinus maenas, was purified to a specific activity of 240 units/mg of protein in the assay conditions described. 2. In one method of purification the enzymic activity could be resolved into two fractions after chromatography on DEAE-cellulose. Fructose 1, 6-diphosphate was able to effect the conversion of one form (peak 1) into the second (peak 2). 3. In the presence of a saturating concentration of fructose 1, 6-diphosphate both forms of the enzyme were kinetically similar. 4. Polyacrylamide-gel electrophoresis of the enzyme 1 day after preparation showed a single protein band. On storage at least three protein bands became visible, all of which were associated with pyruvate kinase activity. 5. Chromatography of the enzyme on Sephadex G-200 indicated a mol.wt. of 247000, but in the presence of fructose 1, 6-diphosphate the elution volume of the enzyme increased corresponding to a mol.wt. of 193000. 6 Dissociation of the enzyme in sodium dodecyl sulphate and 2-mercaptoethanol followed by polyacrylamide-gel electrophoresis produced one major protein band with a mol.wt. of 55000.

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