ULTRACENTRIFUGAL ANALYSIS OF YOLK PROTEINS IN RAINBOW TROUT EGG AND THEIR CHANGES DURING DEVELOPMENT

1965 ◽  
Vol 43 (3) ◽  
pp. 373-379 ◽  
Author(s):  
Kazuo Ando
Keyword(s):  
Molecular Weight ◽  
Rainbow Trout ◽  
Phosphorus Content ◽  
Sedimentation Coefficient ◽  
Egg Yolk ◽  
Yolk Proteins ◽  
Hen’S Egg ◽  
Synchronous Development ◽  
Protein Components ◽  
Ultracentrifugal Analysis

Yolk proteins in Salmo irideus (rainbow trout) eggs were studied by means of ultracentrifugal analysis, and the following facts were clarified. Unfertilized eggs contain two protein components, designated as component I (90% in relative content) and component II (10%). The sedimentation constant for component I is 9.4 S and its molecular weight is approximately 240,000 ~ 260,000. The phosphorus and lipid contents of this major component are similar to those of a lipovitellin in hen's egg yolk, but the molecular weight is considerably smaller than that of the hen's lipovitellin. Component I is split by alkali into two subunits. The sedimentation coefficient for the subunits is 4.9 S and the molecular weight is approximately 120,000. The sedimentation coefficient for component II is 3.1 S, and the phosphorus content is higher than that of component I but is lower than that of hen's phosvitin. A new component of 11.2 S appears at the beginning of the eyed stage, and is inferred to be a protein in the blood formed at this stage. The relative changes of these three components during the synchronous development of embryo from fertilization to the swim-up fry stage were followed.

ISOLATION AND COMPOSITION OF AVIAN EGG YOLK GRANULES AND THEIR CONSTITUENT α- AND β-LIPOVITELLINS

1961 ◽  
Vol 39 (8) ◽  
pp. 1295-1307 ◽  
Author(s):  
R. W. Burley ◽  
W. H. Cook
Keyword(s):  
Molecular Weight ◽  
Low Temperature ◽  
Phosphorus Content ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Egg Yolk ◽  
Low Density ◽  
High Molecular Weight Complex ◽  
Yolk Granules ◽  
Hen’S Egg

The yolk granules from hen's egg represent on a dry basis 23% of the yolk solids, and they contain about 90% of the protein phosphorus, 95% of the iron, and nearly 70% of the calcium in yolk. Ultracentrifugal and other analyses on solutions of the granules show that they are 70% α- and β-lipovitellin in an approximate ratio of 1:1.8, 16% phosvitin, and 12% low-density lipoprotein. The properties and composition of the two lipovitellins isolated from the granules are the same as those isolated from solutions of whole yolk. Further purification reduces the protein phosphorus in α-lipovitellin to 0.50% and in β-lipovitellin to 0.27%, and this confirms that α-vitellin has a higher phosphorus content. Experiments at low temperature suggest that phosvitin exists in the granules as a high molecular weight complex.

FRACTIONATION AND DISSOCIATION OF THE AVIAN LIPOVITELLINS AND THEIR INTERACTION WITH PHOSVITIN

1964 ◽  
Vol 42 (3) ◽  
pp. 395-406 ◽  
Author(s):  
M. W. Radomski ◽  
W. H. Cook
Keyword(s):  
Ionic Strength ◽  
Phosphate Buffer ◽  
Structural Changes ◽  
Phosphorus Content ◽  
Egg Yolk ◽  
Gradient Elution ◽  
Progressive Change ◽  
Hen’S Egg ◽  
Dowex 1 ◽  
Elution Chromatography

Phosvitin and lipovitellin, the granule proteins of hen's egg yolk, were clearly separated by gradient elution on Dowex-1 columns. No phosvitin could be detected in the lipovitellin fraction but the first eluates of the phosvitin fraction contained lipovitellin of high protein phosphorus content. These initial eluates contained only two components sedimenting at rates slightly higher than dimer and monomer lipovitellin. As the lipovitellin monomer does not ordinarily occur in neutral solvents, the slower sedimenting material is either a new component or the monomer stabilized through interaction with phosvitin.Gradient elution chromatography of the total lipovitellins on hydroxyapatite showed that β-lipovitellin was completely eluted by 0.6 M phosphate buffer at pH 6.8 and appeared to be homogeneous. However, α-lipovitellin was heterogeneous: it was eluted over a concentration range of 0.7 to 1.4 M and the protein phosphorus content and dissociative behavior of successive fractions showed a progressive change with increasing ionic strength. Superimposed on this general heterogeneity of α-lipovitellin, there was consistent evidence of two poorly defined components, and three when the α-fraction was rechromatographed. Following dissociation and reassociation, there was no evidence of hybridization between monomers of α- and β-lipovitellin. Changes in the chromatographic patterns of α-lipovitellin following dissociation may indicate hybridization of different α-monomers, but these could also arise from structural changes in the monomers.

SEPARATION AND CHARACTERIZATION OF LIPOVITELLIN FROM HEN EGG YOLK

1958 ◽  
Vol 36 (4) ◽  
pp. 389-398 ◽  
Author(s):  
F. J. Joubert ◽  
W. H. Cook
Keyword(s):  
Molecular Weight ◽  
Phosphorus Content ◽  
Egg Yolk ◽  
Sodium Chloride Solutions ◽  
The Past ◽  
Buffer Solutions ◽  
Separate Protein ◽  
Hen Egg Yolk ◽  
Hen Egg

The major sedimenting fraction of egg yolk, which appears to be homogeneous in sodium chloride solutions, has been fractionated into lipovitellin, phosvitin, and γ-livetin. By dissolving egg yolk in 0.4 M magnesium sulphate and diluting to 0.2 M much of the phosvitin is precipitated and further dilution of the supernatant yields lipovitellin, from which most of the γ-livetin and contaminating phosvitin can be removed by further treatment. Separation and recovery of both lipovitellin and phosvitin by this procedure indicate that phosvitin is a separate protein and not an integral part of the lipovitellin molecule. When the sedimenting fraction in egg yolk is dissolved in buffer solutions at pH 9.0, γ-livetin can be resolved ultracentrifugally from the rest of this fraction, but precipitation by dilution, with ammonium sulphate, or with ethanol, failed to separate the components. Evidently lipovitellin as prepared in the past by similar methods has been a mixture of three proteins. Lipovitellin containing about 10% γ-livetin and 20% lipid had a molecular weight of 3.2 × 105 and a phosphorus content of 0.49%, about half that previously reported.

scholarly journals Two Low-molecular-weight Apoproteins (Apovitellenins I and II) from a Lipoprotein of Goose?s Egg Yolk: A Comparison with Related Species

1982 ◽  
Vol 35 (3) ◽  
pp. 263 ◽  
Author(s):  
AS Inglis ◽  
PM Strike ◽  
RW Burley
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Gel Filtration ◽  
Egg Yolk ◽  
Avian Species ◽  
Pekin Duck ◽  
Muscovy Duck ◽  
Hen’S Egg ◽  
Avian Family

As part of a comparative study of egg yolk from different avian species, the major lipoprotein and its mixed apoproteins from the egg yolk of the chinese goose (Anser cygnoides) have been prepared. From the apoprotein mixture, two new proteins, of molecular weight approximately 10000 and 22000 according to gel electrophoresis in detergent, have been isolated by gel-filtration chromato-graphy in urea. The protein of lower molecular weight corresponds in amino acid sequence to apovitellenin I, a protein previously isolated from other avian species. As a comparison with other members of the same avian family (Anatidae), the amino acid sequence of apovitellenin I from the pekin duck (Anas platyrhynchos) was re-investigated and that of the muscovy duck (Cairina moschata) investigated. These were found to be identical to the sequence of goose's apovitellenin I. The second new protein is similar in composition, molecular weight, and solubility to apovitellenin II, a protein present in small amount in hen's egg yolk. A protein corresponding to apovitellenin II could not, however, be detected in the egg yolk of either species of duck.

scholarly journals Galactothermin, a reversibly heat-precipitable protein of human milk at neutral pH

1970 ◽  
Vol 118 (1) ◽  
pp. 181-186 ◽  
Author(s):  
A. L. Schade ◽  
R. W. Reinhart
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Human Milk ◽  
Skim Milk ◽  
Sedimentation Coefficient ◽  
Amino Acid Residues ◽  
Isolation And Purification ◽  
Neutral Ph ◽  
Polar Species ◽  
Ultracentrifugal Analysis

1. A protein, aggregating at body temperature and solubilizing when cooled, was isolated from fresh human milk at neutral pH and studied for some of its physical, chemical and immunological properties. The name `galactothermin' is proposed for this protein. 2. Isolation and purification of galactothermin involved casein removal from skim milk at pH4.64 followed by centrifugal fractionation of residual protein-containing solutions repeatedly heated and cooled between 40°C and 0°C at pH7.3. 3. The molecular weight by ultracentrifugal analysis and the minimum molecular weight by sum of amino acid residues were 11400 and 14000 respectively. The sedimentation coefficient s25,w was 1.05S and the diffusion coefficient was 7.15×10−7. Reversible aggregation is favoured by increase in protein concentration, ionic strength, temperature, time and approach to the isoionic point of 7.27 from either acidic or alkaline conditions. 4. Among the amino acid residues, proline predominates and non-polar species account for two-thirds of the total. Cysteine and cystine are absent. Analysis of galactothermin showed it to be essentially free of hexose, sialic acid, calcium and phosphate. 5. Galactothermin is antigenic in the rabbit as evidenced by the passive cutaneous anaphylaxis test. Single precipitin lines are produced in immunodiffusion tests. 6. By electrophoresis in polyacrylamide gel at pH4.0 only one sharp band is produced.

scholarly journals The isolation and composition of two phosphoproteins from hen's egg

1970 ◽  
Vol 118 (3) ◽  
pp. 537-542 ◽  
Author(s):  
R. C. Clark
Keyword(s):  
Molecular Weight ◽  
Minor Component ◽  
Egg Yolk ◽  
Terminal Residue ◽  
Hen’S Egg

1. Phosvitin extracted from domestic hen's-egg yolk was resolved on Sephadex G-100 into two phosphoprotein components. 2. The major component has a molecular weight of about 3.4×104 and alanine as an N-terminal residue. Glucosamine is present, but tyrosine is virtually absent. 3. The minor component has a molecular weight of about 2.8×104 and lysine as an N-terminal residue. Missing residues are glucosamine, methionine and leucine. Lysine, histidine, threonine, glycine, phenylalanine and tyrosine contents differ significantly from those of the major component. 4. Sephadex G-100 also removes small amounts of an impurity with a much higher molecular weight.

SEPARATION AND CHARACTERIZATION OF LIPOVITELLIN FROM HEN EGG YOLK

1958 ◽  
Vol 36 (1) ◽  
pp. 389-398 ◽  
Author(s):  
F. J. Joubert ◽  
W. H. Cook
Keyword(s):  
Molecular Weight ◽  
Phosphorus Content ◽  
Egg Yolk ◽  
Sodium Chloride Solutions ◽  
The Past ◽  
Buffer Solutions ◽  
Separate Protein ◽  
Hen Egg Yolk ◽  
Hen Egg

The major sedimenting fraction of egg yolk, which appears to be homogeneous in sodium chloride solutions, has been fractionated into lipovitellin, phosvitin, and γ-livetin. By dissolving egg yolk in 0.4 M magnesium sulphate and diluting to 0.2 M much of the phosvitin is precipitated and further dilution of the supernatant yields lipovitellin, from which most of the γ-livetin and contaminating phosvitin can be removed by further treatment. Separation and recovery of both lipovitellin and phosvitin by this procedure indicate that phosvitin is a separate protein and not an integral part of the lipovitellin molecule. When the sedimenting fraction in egg yolk is dissolved in buffer solutions at pH 9.0, γ-livetin can be resolved ultracentrifugally from the rest of this fraction, but precipitation by dilution, with ammonium sulphate, or with ethanol, failed to separate the components. Evidently lipovitellin as prepared in the past by similar methods has been a mixture of three proteins. Lipovitellin containing about 10% γ-livetin and 20% lipid had a molecular weight of 3.2 × 105 and a phosphorus content of 0.49%, about half that previously reported.

A glyco-phosphoprotein in human milk

1987 ◽  
Vol 54 (2) ◽  
pp. 199-205 ◽  
Author(s):  
Norihiro Azuma ◽  
Kunio Yamauchi
Keyword(s):  
Molecular Weight ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Gel Electrophoresis ◽  
High Performance ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sedimentation Coefficient ◽  
Reversed Phase ◽  
Ultracentrifugal Analysis

SummaryA highly glycosylated phosphoprotein (HGPP) was isolated from a human casein fraction by reversed-phase high-performance liquid chromatography. This component contained carbohydrates to ∼ 38·2% (w/w) and phosphorus to ∼ 1·6% (w/w). The molecular weight of this HGPP as estimated by sodium dodecyl sulphate–polyacrylamide gel electrophoresis ∼ 41000. Ultracentrifugal analysis revealed that the sedimentation coefficient of the HGPP was 2·6S in a 10 mM-imidazole-HCl buffer at pH 7·0 and 27 °C, but this component interacted with human κ-casein and formed a complex with s = 10·4S.

Sign in / Sign up

Export Citation Format

Share Document