The preparation of ribosomal protein from Escherichia coli with lithium chloride and urea

Spitnik‐Elson, Pnina

doi:10.1016/0006-291x(65)90790-4

Cited by 164 publications

(15 citation statements)

References 7 publications

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“…They were resuspended in 4 M LiCl and 6 M urea [19] at 4 "C for 36 h. Precipitated RNAs were removed by centrifugation (27000 x g, 15 min). The supernat,ant containing the ribosomal proteins was dialyzed 4 times against 3 1 0.01 N HC1, then centrifuged to sediment some proteins that were still aggregated.…”

Section: Methodsmentioning

confidence: 99%

Eukaryotic Ribosomal Proteins

Delaunay¹,

Mathieu²,

Schapira³

1972

European Journal of Biochemistry

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The degree of similarity of ribosomal proteins from various species or from various organs within a given species is not clearly known. Using two‐dimensional polyacrylamide‐gel electrophoresis, we have compared liver ribosomal proteins from adult chicken, adult rabbit, fetal and adult calf and ewe. In the rabbit, we also studied ribosomal proteins from kidney, caecal appendix and reticulocytes. In all the cases which we investigated, more than 60 strongly basic proteins are visible. For one given organ, ribosomal proteins do not vary according to the stage of differentiation. Within one given species, there exists few differences from one tissue to another. From one species to another, there are also few differences but the unexpected resemblance of most of the proteins is remarkable.

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Section: Methodsmentioning

confidence: 99%

Eukaryotic Ribosomal Proteins

Delaunay¹,

Mathieu²,

Schapira³

1972

European Journal of Biochemistry

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“…For SDS-gel electrophoretic analysis, ribosomal proteins were extracted according to the method of Spitnik-Elson [22]. Electrophoresis was performed in 5% gels for 8 hr at 8 mA/gel; all solutions contained 6 M urea, all other conditions of the method of Weber and Osborn [23] were maintained, fl-Galactosidase (Boehringer) and BSA (Merck) were used as reference proteins.…”

Section: Methodsmentioning

confidence: 99%

Cross‐linking of ADP‐ribosylated human translocation factor to ribosomes

Üçer

Bermek

1974

FEBS Letters

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“…Ribosomal proteins were extracted according to the method of Spitnik-Elson [17]. 80-S ribosomes corresponding to 100 ,4260 units, 60-S ribosomal subunits corresponding to 60 A,,, units or 40-S ribosomal subunits corresponding to 40 ,4260 units were suspended in 75 pl of medium A minus sucrose (50 mM Tris-HC1 pH 7.4,5 mM MgC12, 25 mM KCl, and 7 mM 2-mercaptoethanol) in 2-ml tubes and mixed with an equal volume of 4 M LiCl in 8 M urea.…”

Section: Preparations Of Ribosomes and Extraction Of Ribosomal Proteinsmentioning

confidence: 99%

Separation of the Proteins in Human Tonsillar Cytoplasmic Ribosomes by Two‐Dimensional Polyacrylamide‐Gel Electrophoresis

Üçler

Bermek

1974

European Journal of Biochemistry

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Proteins of cytoplasmic ribosomes from human tonsillar lymphatic tissue were analyzed by twodimensional polyacrylamide-gel electrophoresis. 34 proteins were found to be associated with the 404 subunit and 35 proteins with the 60-S subunit. 65 proteins had different electrophoretic mobilities. Two proteins found in the small subunit (SIO and S20) co-migrated with two large subunit proteins (L13 and L20 respectively). A comparison of our results with those obtained by the same method for ribosomal proteins from human placenta, rat liver and rabbit reticulocytes suggested a possible identity of the electrophoretic mobilities of 21 -24 40-S subunit proteins and 25-26 6 0 4 subunit proteins.Proteins of cytoplasmic ribosomes from various eucaryotic species and within the same species from various tissues have recently been analyzed by twodimensional gel electrophoresis [l-131. These studies revealed a considerable similarity between the eucaryotic ribosomal proteins of different species. However, some interspecific and also intraspecific differences have been observed [5 -7,111. This communication will report the separation of ribosomal proteins from human tonsillar lymphatic tissue using the method of Kaltschmidt and Wittman MATERIALS AND METHODS ChemicalsAcrylamide, methylene-N,W-bisacrylamide were obtained from Biomol (Ilvesheim) ; urea, trizma base from Baker (Deventer); N,N,N',N'-tetramethylethylendiamine from Serva (Heidelberg). All the other reagent-grade chemicals were purchased from Merck (Darmstadt). Preparations of Ribosomes and Extraction of Ribosomal ProteinsRibosomes [15] and ribosomal subunits [16] from human tonsillar lymphatic tissue were prepared as previously described. Ribosomal proteins were extracted according to the method of Spitnik-Elson [17]. 80-S ribosomes corresponding to 100 ,4260 units, 60-S ribosomal subunits corresponding to 60 A,,, units or 40-S ribosomal subunits corresponding to 40 ,4260 units were suspended in 75 pl of medium A minus sucrose (50 mM Tris-HC1 pH 7.4,5 mM MgC12, 25 mM KCl, and 7 mM 2-mercaptoethanol) in 2-ml tubes and mixed with an equal volume of 4 M LiCl in 8 M urea. After 24 h at 4 "C the RNA precipitate was pelleted by centrifugation for 10 min at 27000 x g in a Sorvall SS-34 rotor. The pellet was resuspended in 50 pl of medium A minus sucrose and extracted again. The supernatant fractions were combined and dialyzed for 30 h against several changes of sample gel solution ~4 1 . Two-Dimensional Electrophoresis and Numbering of Ribosomal ProteinsTwo-dimensional polyacrylamide-gel electrophoresis was performed according to the method of KaltEur. J. Biochem. 50 (1974)

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The preparation of ribosomal protein from Escherichia coli with lithium chloride and urea

Cited by 164 publications

References 7 publications

Eukaryotic Ribosomal Proteins

Eukaryotic Ribosomal Proteins

Cross‐linking of ADP‐ribosylated human translocation factor to ribosomes

Separation of the Proteins in Human Tonsillar Cytoplasmic Ribosomes by Two‐Dimensional Polyacrylamide‐Gel Electrophoresis

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