scholarly journals Further observations on the polynucleotide-induced stimulation of protein synthesis by cell-free preparations from rabbit reticulocytes

1965 ◽  
Vol 97 (2) ◽  
pp. 595-606 ◽  
Author(s):  
KG Nair ◽  
HRV Arnstein
Keyword(s):  
Protein Synthesis ◽  
Messenger Rna ◽  
Free System ◽  
Cell Free System ◽  
Polyadenylic Acid ◽  
Polyuridylic Acid ◽  
Optimum Response ◽  
Ph Range ◽  
Rabbit Reticulocytes ◽  
Stimulation Of

1. The effect of high-molecular-weight RNA from reticulocyte polyribosomes (messenger RNA) on protein synthesis by subcellular fractions derived from reticulocytes, reported by Arnstein, Cox & Hunt (1964), has now been studied in detail. Optimum response of the cell-free system requires 30-50mm-K(+) and approx. 5mm-Mg(2+) in the pH range 7.4-7.6. 2. RNA stimulates the incorporation into protein of both free amino acids and of aminoacyl residues from s-RNA. Stimulation by either RNA or polyuridylic acid is dependent on a labile factor or enzyme, which is present in the; pH5 fraction' and may be concerned with the formation of new polysomes. Quantitatively the response of the cell-free system to RNA is similar to that of polyuridylic acid, and there appears to be competition between messenger RNA and polyuridylic acid or polyadenylic acid.

Gonadotropin stimulation of rat testicular protein synthesis. Polysome isolation and activity in a cell-free system

Biochemistry ◽  
1974 ◽  
Vol 13 (19) ◽  
pp. 3956-3962 ◽  
Author(s):  
Tom O. Abney ◽  
James K. Skipper ◽  
William L. Williams
Keyword(s):  
Protein Synthesis ◽  
Free System ◽  
Cell Free System ◽  
Gonadotropin Stimulation ◽  
A Cell ◽  
Stimulation Of

Proteinsynthese mit Zellextrakten von Micrococcus radiodurans/ Protein Synthesis with Cell Extracts of Micrococcus radiodurans

1978 ◽  
Vol 33 (11-12) ◽  
pp. 948-954
Author(s):  
Annette Widmann ◽  
Roland Süssmuth
Keyword(s):  
Protein Synthesis ◽  
Optimal Conditions ◽  
Free System ◽  
Cell Free System ◽  
E Coli ◽  
Cell Extracts ◽  
Polyuridylic Acid ◽  
A Cell ◽  
Ribosomal Activity ◽  
Cell Free Protein Synthesis

Abstract Pure active ribosomes of cells of Micrococcus radiodurans could be obtained when cultivated in trypton, glucose and nutrient broth by adding natrium citrate. The optimal conditions for a cell-free protein synthesis were investigated at the (polyuridylic acid) dependent polyphenylalanine synthesis. When exchanging ribosomes and S100-fractions with the corresponding fractions of E. coli, we found that the enzyme fractions of M. radiodurans extremely inhibit the ribosomal activity. The incorporation rates in the cell-free system of M. radiodurans yield, at com parable conditions, in relation to E. coli under 10%.

Synthesis of Albumin with Exogenous Mouse-Liver Messenger RNA in a Homologous Cell-Free System

1974 ◽  
Vol 52 (5) ◽  
pp. 429-432 ◽  
Author(s):  
A. J. Faber ◽  
S. H. Miall ◽  
T. Tamaoki
Keyword(s):  
Protein Synthesis ◽  
Total Protein ◽  
Gel Electrophoresis ◽  
Messenger Rna ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Free System ◽  
Cell Free System ◽  
Membrane Bound

RNA extracted from total and membrane-bound polysomes of mouse liver was capable of directing protein synthesis in a homologous cell-free system in the presence of a 0.5 M KCl ribosomal wash fraction. Analysis of the products by polyacrylamide gel electrophoresis and immunoprecipitation showed that newly formed albumin could account for up to 8% of the total protein synthesized.

STIMULATING EFFECT OF DIBUTYRYL CYCLIC ADENOSINE 3′, 5′ MONOPHOSPHATE ON PROTEIN SYNTHESIS IN A THYROIDAL CELL-FREE SYSTEM AT LOW POTASSIUM AND SODIUM CONCENTRATIONS

1973 ◽  
Vol 74 (4) ◽  
pp. 711-722 ◽  
Author(s):  
Gustav Wägar
Keyword(s):  
Protein Synthesis ◽  
Optimal Concentration ◽  
Leucine Incorporation ◽  
Free System ◽  
Cell Free System ◽  
Incubation System ◽  
Low Potassium ◽  
K Concentration ◽  
Stimulation Of ◽  
Precipitable Protein

ABSTRACT The aim of the present study was to investigate the effect of dibutyryl cyclic adenosine 3′, 5′ monophosphate (db-cAMP) on the L-14C-leucine incorporation into TCA-precipitable protein in a thyroidal microsomal cell-free system at low cation concentrations. The basic incubation system contained the following cation concentrations: 18 mm K+, 2.5 mm Na+ and 6 mm Mg2+. Db-cAMP caused a linear stimulation of leucine incorporation in concentrations from 0.5 to 2.5 mm. When the K+ concentration was increased from 18 to 72 mm the stimulating effect of 2.5 mm db-cAMP decreased from 48 to 6 % while the basic incorporation level increased simultaneously. An increase in Na+ from 2.5 to 42.5 mm caused a similar incorporation curve. The optimal concentration of Mg2+ was 6 mm at which concentration the enhancement of leucine incorporation by db-cAMP was most pronounced. Low Ca2+ concentrations did not affect the db-cAMP response. At concentrations over 1 mm, Ca2+ inhibited the protein synthesis without lowering the relative stimulation caused by db-cAMP.

Initiation in einem Polyribosomen-abhängigen Protein-synthetisierenden zellfreien System aus Saccharomyces / Initiation in a Polyribosome-Dependent Protein-Synthesizing Cell-Free System from Saccharomyces

1976 ◽  
Vol 31 (3-4) ◽  
pp. 169-173 ◽  
Author(s):  
Bernd Schulz-Harder ◽  
Ernst-Randolf Lochmann
Keyword(s):  
Protein Synthesis ◽  
Free System ◽  
Cell Free System ◽  
Aurintricarboxylic Acid ◽  
Dependent Protein ◽  
A Cell

Abstract A method to prepare polyribosomes from yeasts by using the french-press is described. The highest yield of polyribosomes was derived from late log-phase cells. These polyribosomes, incubated in a cell-free system, were able to reinitiate protein synthesis, which was shown by inhibiting aminoacid incorporation by aurintricarboxylic acid, edeine and sodiumfluoride. We developed the translational system in order to look for the optimal ion-conditions of a DNA-dependent protein-synthesizing system. We found out that at the optimal MgCL2-concentration (6 mᴍ) protein synthesis was strongly inhibited by Mangan ions which are required for transcription in yeast. If protein-synthesis was carried out with 2 mᴍ and 3 mᴍ MgCl2 maximal aminoacid incorporation was observed at 2 mᴍ and 1.5 mᴍ MnCl2.

scholarly journals Cell-Free Protein Synthesis by Diversifying Bacterial Transcription Machinery

BioTech ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 24
Author(s):  
Marina Snapyan ◽  
Sylvain Robin ◽  
Garabet Yeretssian ◽  
Michèle Lecocq ◽  
Frédéric Marc ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Genomic Sequence ◽  
Synthetic Activity ◽  
Translation System ◽  
Free System ◽  
Transcription Terminator ◽  
Cell Free System ◽  
E Coli ◽  
Bacterial Transcription ◽  
A Cell

We have evaluated several approaches to increase protein synthesis in a cell-free coupled bacterial transcription and translation system. A strong pargC promoter, originally isolated from a moderate thermophilic bacterium Geobacillus stearothermophilus, was used to improve the performance of a cell-free system in extracts of Escherichia coli BL21 (DE3). A stimulating effect on protein synthesis was detected with extracts prepared from recombinant cells, in which the E. coli RNA polymerase subunits α, β, β’ and ω are simultaneously coexpressed. Appending a 3′ UTR genomic sequence and a T7 transcription terminator to the protein-coding region also improves the synthetic activity of some genes from linear DNA. The E. coli BL21 (DE3) rna::Tn10 mutant deficient in a periplasmic RNase I was constructed. The mutant cell-free extract increases by up to four-fold the expression of bacterial and human genes mediated from both bacterial pargC and phage pT7 promoters. By contrast, the RNase E deficiency does not affect the cell-free expression of the same genes. The regulatory proteins of the extremophilic bacterium Thermotoga, synthesized in a cell-free system, can provide the binding capacity to target DNA regions. The advantageous characteristics of cell-free systems described open attractive opportunities for high-throughput screening assays.

scholarly journals Temperature dependency of protein synthesis by cell-free system constructed with polysomes from rice radicle.

1987 ◽  
Vol 56 (1) ◽  
pp. 44-50 ◽  
Author(s):  
Junichi TAKANASHI ◽  
Sachio MARUYAMA ◽  
Nobuyuki KABAKI ◽  
Koichi TAJIMA
Keyword(s):  
Protein Synthesis ◽  
Temperature Dependency ◽  
Free System ◽  
Cell Free System
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