Purification and some properties of Escherichia coli tRNA nucleotidyltransferase. 1977

P Schofield, and K R Williams

Adenylyl (cytidylyl)-tRNA nucleotidyltransferase (ATP (CTP): tRNA adenylyl (cytidylyl)transferase, EC2.7.7.25) has been purified 11,800-fold from a crude extract of Escherichia coli B in an overall yield of 23%. The key step in this purification is the use of a tRNA-Sepharose affinity column. The purified enzyme has a specific activity of approximately 280 mumol of AMP incorporated/min/mg of protein at 37 degrees and has a molecular weight of 52,000 as determined by sodium dodecyl sulfate gel electrophoresis of Sephadex chromatography. The turnover number of the pure enzyme, under optimal assay conditions, is estimated as 21,000, and we believe it constitutes only o.oo6% of the total cellular protein. Both AMP- and CMP-incorporating activities have an identical isoelectric point of 5.85. The AMP-incorporating activity of the enzyme is inhibitied by some transition metal chelating agents but not by others.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D012316 RNA Nucleotidyltransferases Enzymes that catalyze the template-directed incorporation of ribonucleotides into an RNA chain. EC 2.7.7.-. Nucleotidyltransferases, RNA
D012343 RNA, Transfer The small RNA molecules, 73-80 nucleotides long, that function during translation (TRANSLATION, GENETIC) to align AMINO ACIDS at the RIBOSOMES in a sequence determined by the mRNA (RNA, MESSENGER). There are about 30 different transfer RNAs. Each recognizes a specific CODON set on the mRNA through its own ANTICODON and as aminoacyl tRNAs (RNA, TRANSFER, AMINO ACYL), each carries a specific amino acid to the ribosome to add to the elongating peptide chains. Suppressor Transfer RNA,Transfer RNA,tRNA,RNA, Transfer, Suppressor,Transfer RNA, Suppressor,RNA, Suppressor Transfer

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