Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8.

Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Research Abstract Details 

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Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Abstract Text:

M Hara-Yokoyama,S Yokoyama,T Miyazawa,

Glutamyl-tRNA synthetase has been isolated from an extreme thermophile, Thermus thermophilus HB8. The enzyme has been purified to homogeneity by successive chromatography on columns of DEAE-cellulose, DEAE-Sephacel, phosphocellulose and hydroxyapatite. 11.7 mg of purified enzyme has been obtained from 2 kg of T. thermophilus cells, with a purification factor of 600 with an 11% yield. From gel permeation chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis, the enzyme is found to be a monomer protein with a molecular weight of 50,000. The optimum temperature for the aminoacylation of T. thermophilus tRNAGlu is 65 degrees C, and the optimum pH range is 8.0-9.0, in the presence of 5 mM Mg2+. The Km values for ATP, L-glutamate, and T. thermophilus tRNAGlu are 230 microM, 70 microM, and 0.65 microM, respectively, in the presence of 50 mM KCl and 10 mM MgCl2 at pH 8.0 at 65 degrees C. Escherichia coli tRNA2Glu is also a good substrate with a Km value of 0.60 microM at 65 degrees C. The mole fractions of Arg and Leu residues are higher and that of Asx residues is lower than those of E. coli glutamyl-tRNA synthetase. Glutamyl-tRNA synthetase from T. thermophilus is remarkably thermostable; even after incubation for 9 h at 65 degrees C, 70% of the enzyme activity is retained in the absence of any protecting factors. Such an extremely thermostable enzyme with a low molecular weight will be useful for detailed physiochemical analyses on the molecular mechanism of strict recognition by aminoacyl-tRNA synthetases.

Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Publishing Authors By Initials

M Hara-Yokoyama,S Yokoyama,T Miyazawa,

For similar bacteria: gram-negative bacteria: gram-negative aerobic bacteria: gram-negative aerobic rods and cocci: thermus research abstracts see: bacteria: gram-negative bacteria: gram-negative aerobic bacteria: gram-negative aerobic rods and cocci: thermus research

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Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 96

Page Numbers: 1599-607

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Nov

YEAR: 1984

Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Keywords Mesh Terms:

KEYWORDS: Thermus

MESH TERMS: enzymology

Chemical & Substance for Abstract: Purification and characterization of glutamyl-tRNA synthetase from an extreme thermophile, Thermus thermophilus HB8. Information

Substance Name: Glutamate-tRNA Ligase

Registry Number: EC 6.1.1.17

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Country: JAPAN

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MEDLINETA: J Biochem

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