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Biomedical subjects

D P Leader

Publications and source records attributed to D P Leader.

At least 73 records · Page 4Linked to original sources
PubMed1978

Characterization of the acidic phosphorprotein of eukaryotic ribosomes using a new system of two-dimensional gel-electrophoresis.

1. A modified method of two-dimensional gel electrophoresis has been developed for detecting acidic eukaryotic ribosomal proteins. 2. Using this method it has been demonstrated that the major phosphoprotein (Lgamma) of mouse ascites and hamster fibroblast 60-S ribosomal subunits is an acidic protein, apparently analagous to L7/12 of Escherichia coli, and not a neutral protein as previously thought. Electrophoretic resolution of phosphorylated and non-phosphorylated forms of Lgamma has enabled the stoichiometric nature of the phosphorylation (previously deduced from measurement of the specific radioactivity of the ATP pool) to be confirmed. 3. When ascites cells were incubated for 3 h there appeared an altered form of Lgamma which is most likely produced by proteolytic cleavage of the original form. The extent of phosphorylation of Lgamma was decreased by one-half or more in these circumstances. 4. Phosphoprotein Lgamma was found to be almost completely phosphorylated in both polyribosomes and monoribosomes isolated from hamster fibroblasts. Thus the function of the phosphorylation of Lgamma appears not to be concerned with the inactivation of ribosomes.

Animals↗
PubMed1978

The phosphorylation of ribosomal proteins L14 and S3 in Krebs II ascites cells.

Krebs II ascites cells incubated in Earle's saline (lacking glucose and amino acids) contain ribosomes with proteins S6 and Lgamma phosphorylated, as do ascites cells grown in the peritonea of mice or hamster fibroblasts grown in Eagle's medium. When ascites cells were incubated in Eagle's medium (containing glucose and amino acids) there was extensive glycolysis, producing very acidic conditions, and ribosomal proteins S3 and L14 became phosphorylated whereas Lgamma became dephosphorylated. This altered pattern of phosphorylation could not be produced merely by incubating ascites cells in Earle's saline at a decreased pH, but a rather similar pattern was produced when Earle's saline was supplemented with amino acids (but with glucose still omitted). These results suggest that depriving ascites cells of glucose may induce the synthesis of a protein (or proteins), necessary for alteration of the pattern of phosphorylation of the ribosomal proteins.

Cells, Cultured↗
PubMed1975

Enzymic iodination of eukaryotic ribosomal subunits. Characterization and analysis by two-dimensional gel electrophoresis.

1. Conditions are described for the enzymic iodination of ribosomal subunits from rat liver. The reaction is relatively insensitive to broad changes in the concentration of KCl, allowing subunits to be studied under conditions which minimize their dimerization. 2. Mixtures of extracted ribosomal proteins were iodinated with (125)I, the proteins separated by two-dimensional gel electrophoresis and the radioactivity in each protein was determined. Thus 19 out of 23 of the proteins of the small subunit and 25 out of 33 of the proteins of the large subunit were labelled. Iodination should therefore be a suitable method for studying the topography of the ribosomal proteins of rat liver. 3. When the intact 40S subunit (rather than the extracted mixture of proteins) was iodinated, 18 of the 19 proteins were still labelled. However five of these were labelled less strongly than before. When the intact 60S subunit was iodinated, 17 of the 25 proteins were still labelled, although six of these were labelled less strongly. 4. These results show that in rat liver most of the ribosomal proteins of both subunits are at least partially at the surface of the particles. They are also consistent with the idea that the proportion of the ribosomal proteins in the interior of the particle may be greater for the 60S subunit than for the 40S subunit.

Animals↗
PubMed1975

The binding sites for tRNA on eukaryotic ribosomes.

We have studied the non-enzymic binding of phe-tRNA to ribosomes from rat liver using deacylated tRNA to inhibit binding to the P-site and puromycin (5 x 10-minus3M) to inhibit binding to the A-site. We conclude that at a low concentration of magnesium ions (10mM) phe-tRNA is bound only at the A-site of 80S irbosomes, whereas at a high concentration of magnesium ions (40mM) phe-tRNA is also bound at the P-site. Studies with edeine indicate that, during non-enzymic binding of phe-tRNA, eukaryotic ribosomes (in contrast to prokarotic ribosomes) have the A-site of the 60S subunit and the initiation site of the 40S subunit juxtaposed. This may account for the differences observed, in formation of diphenylalanyl-tRNA and phenylalanyl-puromycin, between phe-tRNA bound non-enzymically to the P-sites of eukaryotic and prokaryotic ribosomes.

Animals↗