CHROMATOGRAPHIC SEPARATION OF PHOSVITIN, α- AND β-LIPOVITELLIN OF EGG YOLK GRANULES ON TEAE-CELLULOSE

1964 ◽  
Vol 42 (8) ◽  
pp. 1203-1215 ◽  
Author(s):  
M. W. Radomski ◽  
W. H. Cook
Keyword(s):  
Ionic Strength ◽  
Egg Yolk ◽  
Gradient Elution ◽  
Partial Recovery ◽  
Linear Gradient ◽  
Yolk Granules ◽  
Sedimentation Behavior ◽  
Strength Limit ◽  
The Individual ◽  
Low Ionic Strength

The two components of lipovitellin and the three major components of yolk granules, phosvitin, α- and β-lipovitellin, have been separated by gradient elution chromatography on TEAE-cellulose. A 0.2 M phosphate buffer (pH 6.8) had the necessary ionic strength to dissolve these proteins and when applied in this solvent all components except β-lipovitellin were retained by the column. A linear gradient of ionic strength (limit buffer 0.2 M phosphate plus 0.5 M NaCl) was used to remove the other components. Recovery was essentially complete and the composition and properties of the individual components were similar to those obtained by previous chromatographic methods that gave only partial recovery. An additional component eluted after α-lipovitellin and before phosvitin, previously observed in Dowex-1 separations, was also observed by the present method. The composition, sedimentation behavior, and absorption spectra of this component indicate that it is a soluble complex of phosvitin and lipovitellin. When granules are dissolved in alkaline solvents (pH 9.4) of low ionic strength (0.05), phosvitin is not evident as a separate component during ultracentrifugation, but appears as the ionic strength is increased.

An Evaluation of a Plasma Fractionation System

1960 ◽  
Vol 4 (01) ◽  
pp. 031-044
Author(s):  
George Y. Shinowara ◽  
E. Mary Ruth
Keyword(s):  
Biological Activity ◽  
Ionic Strength ◽  
Plasma Proteins ◽  
High Concentration ◽  
Significant Evidence ◽  
Native Proteins ◽  
Mobility Data ◽  
Fractionation Procedure ◽  
The Individual ◽  
Low Ionic Strength

SummaryFour primary fractions comprising at least 97 per cent of the plasma proteins have been critically appraised for evidence of denaturation arising from a low temperature—low ionic strength fractionation system. The results in addition to those referable to the recovery of mass and biological activity include the following: The high solubilities of these fractions at pH 7.3 and low ionic strengths; the compatibility of the electrophoretic and ultracentrifugal data of the individual fractions with those of the original plasma; and the recovery of hemoglobin, not hematin, in fraction III obtained from specimens contaminated with this pigment. However, the most significant evidence for minimum alterations of native proteins was that the S20, w and the electrophoretic mobility data on the physically recombined fractions were identical to those found on whole plasma.The fractionation procedure examined here quantitatively isolates fibrinogen, prothrombin and antithrombin in primary fractions. Results have been obtained demonstrating its significance in other biological systems. These include the following: The finding of 5 S20, w classes in the 4 primary fractions; the occurrence of more than 90 per cent of the plasma gamma globulins in fraction III; the 98 per cent pure albumin in fraction IV; and, finally, the high concentration of beta lipoproteins in fraction II.

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.

Conformational Transitions in Escherichia coli Ribosomal RNAs as Visualized by Dedicated STEM

1988 ◽  
Vol 46 ◽  
pp. 176-177
Author(s):  
J.S. Wall ◽  
V. Maridiyan ◽  
S. Tumminia ◽  
J. Hairifeld ◽  
M. Boublik
Keyword(s):  
Ionic Strength ◽  
Three Dimensional ◽  
Conformational Transitions ◽  
Dark Field ◽  
Dimensional Structure ◽  
Ribosomal Rnas ◽  
Field Mode ◽  
Freeze Dried ◽  
High Resolution Images ◽  
Low Ionic Strength

The high contrast in the dark-field mode of dedicated STEM, specimen deposition by the wet film technique and low radiation dose (1 e/Å2) at -160°C make it possible to obtain high resolution images of unstained freeze-dried macromolecules with minimal structural distortion. Since the image intensity is directly related to the local projected mass of the specimen it became feasible to determine the molecular mass and mass distribution within individual macromolecules and from these data to calculate the linear density (M/L) and the radii of gyration.2 This parameter (RQ), reflecting the three-dimensional structure of the macromolecular particles in solution, has been applied to monitor the conformational transitions in E. coli 16S and 23S ribosomal RNAs in solutions of various ionic strength.In spite of the differences in mass (550 kD and 1050 kD, respectively), both 16S and 23S RNA appear equally sensitive to changes in buffer conditions. In deionized water or conditions of extremely low ionic strength both appear as filamentous structures (Fig. la and 2a, respectively) possessing a major backbone with protruding branches which are more frequent and more complex in 23S RNA (Fig. 2a).

scholarly journals A novel procedure for the rapid purification of plastocyanin from pea (Pisum sativum) leaves

1981 ◽  
Vol 193 (1) ◽  
pp. 375-378 ◽  
Author(s):  
A R Ashton ◽  
L E Anderson
Keyword(s):  
Ionic Strength ◽  
Pisum Sativum ◽  
Deae Cellulose ◽  
High Concentrations ◽  
Rapid Purification ◽  
Low Ionic Strength

Plastocyanin is soluble at high concentrations (greater than 3 M) of (NH4)2SO4 but under these conditions will adsorb tightly to unsubstituted Sepharose beads. This observation was utilized to purify plastocyanin from pea (Pisum sativum) in two chromatographic steps. Sepharose-bound plastocyanin was eluted with low-ionic-strength buffer and subsequently purified to homogeneity by DEAE-cellulose chromatography.

scholarly journals Crystallization of tuna ferricytochrome c at low ionic strength.

1990 ◽  
Vol 265 (8) ◽  
pp. 4177-4180
Author(s):  
M H Walter ◽  
E M Westbrook ◽  
S Tykodi ◽  
A M Uhm ◽  
E Margoliash
Keyword(s):  
Ionic Strength ◽  
Ferricytochrome C ◽  
Low Ionic Strength

scholarly journals Reaction of Myosin with 1-Fluoro-2,4-dinitrobenzene at Low Ionic Strength

1969 ◽  
Vol 244 (3) ◽  
pp. 648-657 ◽  
Author(s):  
M Bárány ◽  
G Bailin ◽  
K Bárány
Keyword(s):  
Ionic Strength ◽  
Low Ionic Strength
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