Ribosomal proteins from goose reticulocytes are not histones

1968 ◽  
Vol 46 (12) ◽  
pp. 1507-1514 ◽  
Author(s):  
J. M. Neelin ◽  
G. Vidali

Ribosomes were isolated from goose reticulocytes after lysis with saponin in 50 mM KCl, 1.5 mM MgCl2, 1 mM Tris, pH 7.5. Maximum yields of ribosomes were obtained about 4 days after injection of the birds with Phenylhydrazine, but ribosomal proteins did not vary with the stage of recovery.Ribosomal proteins (extracted with either HCl–urea or LiCl–urea) differed generally from histones (extracted with either HCl or HCl–urea) according to amino acid composition and to electrophoretic patterns in starch gel and in Polyacrylamide gel, but a few zones of ribosomal protein appeared to coincide electrophoretically with the main histone components. Since all of the former proteins were eluted unretarded from Amberlite CG-50 in 9% guanidinium chloride, in which all histones are adsorbed, we conclude that histones and ribosomal proteins are different classes of protein.The hazards of assuming chemical identities of proteins on the basis of limited electrophoretic evidence and the risks of misleading cross-contamination of cell fractions were demonstrated.

1964 ◽  
Vol 17 (4) ◽  
pp. 1001 ◽  
Author(s):  
JT Bellair ◽  
OM Mauritzen

Crude IX-histone, obtained from the original histone complex by precipitation of the f3-and y-histones with ethanol, has been shown by starch-gel electrophoresis to contain 13 components. The fractionation of IX-histone by exclusion chromatography on Sephadex G-200 is reported. While none of these components have been obtained pure in the present study, considerable purification of components 2, 3, 4, 5, 8, 9, 10, and 11 has been achieved, and their amino acid composition leaves no doubt that o::-histones represent a muoh larger family of "lysine-rich" proteins than was hitherto supposed.


1966 ◽  
Vol 44 (8) ◽  
pp. 1221-1233 ◽  
Author(s):  
A. M. MacQuillan ◽  
S. T. Bayley

The results of several procedures for solubilizing protein from pea-seedling ribosomes are described. Pancreatic ribonuclease (RNAase), alone at pH 8.0 and low ionic strength, solubilized 10%, whereas subsequent treatment with dilute HC1 solubilized a further 29%. RNAase in 8 M urea at pH 5.5 solubilized more than 90% of the ribosomal protein. After removal of ribonucleic acid fragments, all of this material, termed "total protein", remained in solution in 0.05 M acetate buffer at pH 5. Proteins solubilized by these three procedures were subjected to electrophoresis on starch–urea gels, amino acid analyses, and fingerprinting after digestion with trypsin and chymotrypsin. The protein soluble in RNAase probably contains only one or two components and is distinct in amino acid and peptide composition from the remainder of the ribosomal protein. Protein soluble in HC1 is representative of much of the "total protein" and both HC1-soluble and "total protein" are complex. Fingerprints showed that although RNAase-extracted protein and three fractions of "total protein" obtained by gel electrophoresis were different, they also possessed a number of peptides in common. It was concluded that the number of components in pea-ribosomal proteins is probably of the order of 16, the number of bands seen in gel-electrophoretic patterns.


1965 ◽  
Vol 18 (6) ◽  
pp. 1207 ◽  
Author(s):  
EOP Thompson ◽  
IJ O'donnell

Starch-gel electrophoresis of wool proteins extracted from reduced and carboxymethylated wool gives a complex pattern in which there are two major protein bands. By a combination of chromatography on DEAE-cellulose and gelfiltration in buffers containing 8M urea these two protein components have been isolated. The amino acid composition and some properties of these two fractions are reported. A comparison of the amino acid composition and of peptide maps of tryptic digests of the two fractions shows distinct differences between them, and by labelling with 2-[14C]iodoacetate the distribution of the peptides containing S-carboxymethylcysteine residues were also shown to be different.


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