Isolation of κ-casein-like fractions from human whole casein by chromatography on thiol-Sepharose, hydroxyapatite and DEAE-cellulose

1981 ◽  
Vol 48 (3) ◽  
pp. 429-436 ◽  
Author(s):  
Jean-Marc Chobert ◽  
Francesco Addeo ◽  
Bruno Ribadeau Dumas
Keyword(s):  
Affinity Chromatography ◽  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Phosphate Buffer ◽  
Deae Cellulose ◽  
The Other ◽  
Starch Gel Electrophoresis ◽  
Sh Groups ◽  
Starch Gel

SummaryDifferent fractions of human κ-casein characterized by their sensitivity to chymosin were separated, after reduction, by affinity chromatography on thiol-Sepharose in a 0·1 m-Tris-HCl buffer, pH 7·0, containing 7 M-urea, 0·3 m-NaCl and 1 mm-EDTA. The fraction retained on the column and bearing the SH-groups was then chromatographed on hydroxyapatite in a 5 mm-phosphate buffer, pH 6·8, containing 0·2 m-KC1, 4·5 m-urea and 2 mm 2-mercaptoethanol. The fraction not retained on this column was purified by chromatography on DEAE-cellulose in a 6·6 m-urea, 0·02 m-imidazole, 0·02 m 2-mercaptoethanol buffer, pH 7·0. Two chymosinsensitive fractions were identified by starch-gel electrophoresis at pH 8·6; one of low electrophoretic mobility, the other a smeared band covering the whole distance of migration, in which several bands with mobilities greater than those of β-caseins could be detected.

scholarly journals A chromatographic preparation of ox growth hormone

1966 ◽  
Vol 100 (3) ◽  
pp. 593-600 ◽  
Author(s):  
M Wallis ◽  
HBF Dixon
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Gel Electrophoresis ◽  
Anterior Pituitary ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Highly Active ◽  
Cross Linkage ◽  
Starch Gel

1. A method is described for the chromatographic preparation of ox growth hormone. It involves chromatography of an extract of anterior pituitary lobes on DEAE-cellulose, followed by rechromatography on a dextran gel of low cross-linkage (Sephadex G-100). 2. The product is highly active in growth-hormone assays, and is obtained in good yield. It was homogeneous by several criteria, but showed some heterogeneity on starch-gel electrophoresis. 3. The molecular weight of the hormone was estimated from its behaviour on gel-filtration columns under various conditions. Evidence that the hormone may dissociate into sub-units under some conditions is presented.

scholarly journals ENZYMATICALLY PRODUCED SUBUNITS OF PROTEINS FORMED BY PLASMA CELLS IN MICE

1962 ◽  
Vol 115 (3) ◽  
pp. 623-639 ◽  
Author(s):  
J. L. Fahey ◽  
Brigitte A. Askonas
Keyword(s):  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Gamma Globulin ◽  
Immunological Methods ◽  
Antigenic Determinants ◽  
Antibody Activity ◽  
Starch Gel Electrophoresis ◽  
Myeloma Protein ◽  
Γ Globulins ◽  
Starch Gel

Gamma globulin and antibody obtained from inbred C3H mice are split by papain and cysteine into fragments roughly one-third the size of the original Molecule (S20,w = 3.5S). The papain digests were characterized by starch gel electrophoresis and immunological methods. The highly heterogeneous fragments could be divided into two groups with distinct antigenic determinants (S and F), which were separated by DEAE ion-exchange cellulose chromatography. Approximately two-thirds of the fragments had S antigenic groupings and one-third had F antigenic groupings. These data are consistent with the view that mouse gamma globulin is split by papain and cysteine into three major fragments, two of which are of the S type and one of the F type. Antibody activity of the original molecule was present in the S fragments. Although the S fragments did not precipitate the antigen (hemocyanin) they were shown to bind antigen specifically in the manner of univalent antibodies. The S fragments of normal γ-globulin were very heterogeneous with a broad spectrum of electrophoretic mobilities. Comparison of S fragments from slow and fast migrating globulins showed that the mobilities of the original γ-globulin samples were largely reflected in the mobilities of their S fragments. Additional observations indicated that the F fragments also may help to determine the electrophoretic mobility of intact γ-globulin molecules. S fragments of differing electrophoretic mobility were shown to have the same antigenic determinants, indicating that the structural differences responsible for the electrophoretic mobility differences were not involved in the antigenic groupings identified with rabbit antisera. The F fragments of normal γ-globulin migrated more rapidly than the S fragments, were less heterogeneous, and showed several bands on starch gel electrophoresis. The F fragments differed antigenically from the S fragments, and had no antibody activity. Two groups of F fragments (F and F') were detected with some antisera. The γ-myeloma protein (5563) formed in a C3H plasma cell tumor and similarly fragmented by treatment with papain and cysteine, produced much more discrete S and F components than were found in the normal γ-globulin digest. The electrophoretic properties of the myeloma protein fragments were within the range observed for normal γ-globulin fragments. Although the γ-myeloma protein shares antigenic determinants with normal γ-globulins it lacks some of the antigenic groupings present in the γ-globulin preparation. Both S and F fragments from the myeloma protein share antigenic determinants with the corresponding fragments from normal γ-globulin. In addition, both S and F fragments of normal γ-globulin possess antigenic groupings not present in fragments of the γ-myeloma protein, accounting for the antigenic deficiency observed on comparison of the γ-myeloma protein with normal γ-globulins.

Identification of Australian barley cultivars by laboratory methods: gel electrophoresis and gel isoelectric focusing of the endosperm proteins

1977 ◽  
Vol 17 (89) ◽  
pp. 1020 ◽  
Author(s):  
J McCausland ◽  
CW Wrigley
Keyword(s):  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Water Soluble ◽  
The Other ◽  
Starch Gel Electrophoresis ◽  
Laboratory Methods ◽  
Barley Cultivars ◽  
Starch Gel ◽  
Electrophoretic Techniques ◽  
Gel Isoelectric Focusing

A range of laboratory methods was examined for their ability to distinguish between 19 barley cultivars currently grown in Australia. Aleurone colour, revealed after mechanical or chemical dehulling, differentiated Abyssinian, Atlas, Cape and Corvette from the other cultivars. Peroxidase and phenol testing were not useful. Seven different patterns were obtained for the hordeins of lowest mobility by starch gel electrophoresis. Further distinction was provided by flat gel isoelectric focusing of the water-soluble and hordein proteins for which 13 different pattern-groupings were obtained. The two electrophoretic techniques complemented one another, so that the use of both methods left only a few cultivars that could not be distinguished.

STUDIES ON PLASMINOGEN: IV. ALTERATION OF BOVINE AND HUMAN PLASMINOGENS DURING ISOLATION

1966 ◽  
Vol 44 (4) ◽  
pp. 469-473 ◽  
Author(s):  
John Y. S. Chan ◽  
Edwin T. Mertz
Keyword(s):  
Ionic Strength ◽  
Gel Electrophoresis ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Major Band ◽  
Starch Gel ◽  
Human Plasminogen

Bovine and human plasminogen preparations were analyzed by starch-gel electrophoresis at pH 2.5 and 0.10 ionic strength. The bands were activated with urokinase and the proteolytic and esterolytic activities measured. Bovine euglobulin contains one plasminogen band, B-1. Plasminogen prepared from bovine euglobulin by continuous electrophoresis at pH 3.5 contains B-1 and a faster plasminogen band, B-2. B-1 and B-2 are also found in bovine plasminogen prepared by DEAE-cellulose chromatography. All three preparations on activation give the same two plasmin bands on starch gel. Human euglobulin also contains two active plasminogen bands H-1 and H-2. Plasminogen prepared from human euglobulin by continuous electrophoresis at pH 3.5 contains H-1, H-2, and a faster minor plasminogen band, H-3. All highly purified human plasminogens derived from Cohn fraction III contain either H-3 as a major band and an additional plasminogen band, H-4, or only H-3, but no H-1 and H-2. On activation with urokinase or streptokinase, human plasminogen preparations give one or two plasmin bands. It is concluded that bovine B-2 and human H-3 and H-4 are altered forms of euglobulin plasminogen created during isolation procedures. Essentially pure human H-3 can be prepared by continuous electrophoresis from Cutter plasminogen.

STUDIES ON THE LIVETINS OF EGG YOLK: III. HETEROGENEITY AND IMMUNOELECTROPHORETIC BEHAVIOR OF BETA-LIVETIN

1966 ◽  
Vol 44 (10) ◽  
pp. 1357-1364 ◽  
Author(s):  
Shun-Fong Hui ◽  
R. H. Common
Keyword(s):  
Serum Albumin ◽  
Gel Electrophoresis ◽  
Egg Yolk ◽  
The Other ◽  
Positive Staining ◽  
Staining Reaction ◽  
Starch Gel Electrophoresis ◽  
Hen’S Egg ◽  
Starch Gel ◽  
The One

Starch-gel electrophoresis of the total livetins of hen's egg yolk resolved 16 zones: seven major zones, six minor zones, and three faint, diffuse zones. One zone was identified with the major component of paper electrophoretic alpha-livetin and hence with serum albumin. Four of the major zones were identified with the major components of paper electrophoretic beta-livetin on the one hand, and with an electrophoretically heterogeneous livetin antigen (livetin antigen 3) on the other hand, thus establishing the electrophoretic heterogeneity and relative immunological homogeneity of the paper electrophoretic beta-livetin fraction. The other two major starch-gel electrophoretic zones were identified as transferrins by their positive staining reaction for iron and comparison of their mobilities with two corresponding serum starch-gel fractions.

Benzylamine oxidase and histaminase: purification and crystallization of an enzyme from pig plasma

1964 ◽  
Vol 161 (983) ◽  
pp. 153-167 ◽  
Keyword(s):  
Ammonium Sulphate ◽  
Gel Electrophoresis ◽  
Copper Content ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Anaerobic Conditions ◽  
Concentrated Solutions ◽  
Radioactivation Analysis ◽  
Starch Gel ◽  
Benzylamine Oxidase

The enzyme benzylamine oxidase of pig plasma has been purified and some of the properties of the pure preparation have been studied. The purification procedure included several precipitations with ammonium sulphate and separations of proteins by column chromatography, first on DEAE-cellulose, followed by DEAE-Sephadex and lastly on a hydroxyapatite column. Crystals were prepared from solutions of the purified enzyme by adding ammonium sulphate. The crystalline preparation was homogeneous when studied by starch-gel electrophoresis and by ultracentrifugation. The molecular weight, as determined on the analytical ultracentrifuge, was 195 000. The copper content of the enzyme, as determined by radioactivation analysis, was about four atoms of Cu per molecule of enzyme. Concentrated solutions of the enzyme had a pink colour; the colour disappeared when substrate (benzylamine) was added under anaerobic conditions. The amines which were tested and found to be oxidized by the pure enzyme were: benzylamine, histamine, mescaline and 4-picolylamine. The affinity of the enzyme for benzylamine was more than one hundred times that for histamine.

A Genetic Comparison of Sympatric Populations of Sand Lance (Genus Ammodytes) from the Region East of Cape Soya, Japan

1989 ◽  
Vol 46 (11) ◽  
pp. 1945-1951 ◽  
Author(s):  
Hiroaki Okamoto
Keyword(s):  
Genetic Structure ◽  
Gel Electrophoresis ◽  
Distinct Species ◽  
The Other ◽  
Starch Gel Electrophoresis ◽  
Sympatric Populations ◽  
Sand Lance ◽  
Protein Coding ◽  
Starch Gel ◽  
Genetic Comparison

Sand fance (Genus Ammodytes) collected from four stations off Japan and one station at Kodiak, Alaska were genetically characterized at 17 protein coding loci using starch-gel electrophoresis. Sand lance in Wakkanai (Cape Soya, Japan) consist of two genetically distinct groups. They are fixed for different alleles at four loci (Ldh-2, -3, G3pdh-2, and Mdhp-2). The genetic structure of one of the groups (Wakkanai-a group, W-a) is similar to that of A. personatus around Japan. The other group (Wakkanai-b group, W-b) has different genetic structure from either A. personatus or the Alaskan collection, which is presumed to belong to A. hexapterus. It is not presently possible to identify the affiliation of the W-b group; however, despite its sympatry with the W-a group, it is reproductively isolated and therefore is probably a distinct species occurring northeast of Hokkaido.

Ontogenetic Variation in the Multiple Hemoglobins of Arctic Charr, Salvelinus alpinus

1989 ◽  
Vol 46 (5) ◽  
pp. 804-809 ◽  
Author(s):  
M. A. Giles ◽  
D. M. Rystephanuk
Keyword(s):  
Life Cycle ◽  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Salvelinus Alpinus ◽  
Arctic Charr ◽  
Ontogenetic Variation ◽  
Starch Gel Electrophoresis ◽  
Free Swimming ◽  
Total Hemoglobin ◽  
Starch Gel

Changes during development in the multiple hemoglobins of hatchery-reared Arctic charr, Salvelinus alpinus, that originated from an anadromous stock from Nauyuk Lake, Northwest Territories were examined by starch gel electrophoresis. A total of 10 anodic and 7 cathodic hemoglobins were expressed during the life cycle of the charr. Embryos and newly-hatched alevins exhibited 10 anodic and 3 cathodic components. Of these embryonic hemoglobins, 5 anodic components and possibly 1 cathodic component with similar electrophoretic mobility were observed in older fish. In free-swimming fry 3–4 new cathodic components were expressed. Two phenotypes (designated as 3-C or 5-C) were identified in postembryonic charr, distinguished by the presence of either 3 or 5 cathodic hemoglobins, respectively. In both phenotypes the proportion of cathodic hemoglobins increased progressively with age to a maximum of 22 and 18% of total hemoglobin in the 3-C and 5-C phenotypes, respectively.

Patterns of evolution and migration in the arctic ground squirrel, Spermophilus parryii (Richardson)

1977 ◽  
Vol 55 (4) ◽  
pp. 748-758 ◽  
Author(s):  
Charles F. Nadler ◽  
Robert S. Hoffmann
Keyword(s):  
Body Size ◽  
Gel Electrophoresis ◽  
Ground Squirrel ◽  
The Other ◽  
The Arctic ◽  
Starch Gel Electrophoresis ◽  
Siberian Population ◽  
Spermophilus Parryii ◽  
Starch Gel ◽  
And Migration

Serum transferrins and 11 other genetically controlled proteins representing 17 loci were examined by starch-gel electrophoresis from Siberian, Alaskan, and Canadian populations of Spermophilus parryii. Six transferrin alleles were identified. Arctic populations (S. p. parryii, S. p. osgoodi) were characterized by Tf 6 occurring alone or together with Tf 7 whereas middle and subarctic populations exhibited Tf 7 occurring either alone (S. p. ablusus, S. p. lyratus, S. p. plesius) or together with Tf 5 (S. p. plesius). Tf 8, Tf 9, and Tf 19 constituted local variants. Tf 6 displayed a clinal distribution, increasing in frequency eastward and paralleling a clinal increase in body size. Three PGM2 alleles were observed, the frequencies of which tend to differentiate arctic S. p. parryii from subarctic S. p. ablusus. G6PD-b occurred uniformly in North America and in one Siberian population; a second population (two specimens) exhibited G6PD-a, thereby suggesting that G6PD polymorphism may be present in Siberian S. parryii. The other nine proteins were monomorphic in all Holarctic populations.

A COMPARISON OF HISTONES FROM CHICKEN TISSUES BY ZONE ELECTROPHORESIS IN STARCH GEL

1961 ◽  
Vol 39 (3) ◽  
pp. 485-491 ◽  
Author(s):  
J. M. Neelin ◽  
G. C. Butler
Keyword(s):  
Cell Differentiation ◽  
Ionic Strength ◽  
Gel Electrophoresis ◽  
The Other ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Patterns ◽  
Zone Electrophoresis ◽  
Starch Gel ◽  
Chicken Erythrocytes ◽  
Characteristic Zone

Histories were extracted at pH 1.7 from washed nuclei of chicken erythrocytes, spleen, liver, and testis and compared by starch-gel electrophoresis at pH 5.0, ionic strength 0.020. Spleen and liver histories displayed the most complex electrophoretic patterns with 18 zones each and differed only in relative proportions of certain zones. Erythrocyte histone contained a characteristic zone while lacking a group present in spleen and liver histones. Testis histone with only seven zones differed markedly from the other three. These results were consistent with chromatograms of erythrocyte, spleen, and liver histones on sodium IRC-50. The suggested correlation of tissue-specific histones with cell differentiation is discussed.

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