Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium

Alfredo Colonna; Gennaro Ciliberto; Rita Santamaria; Filiberto Cimino; Francesco Salvatore

doi:10.1007/BF00224919

Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium

Alfredo Colonna, Gennaro Ciliberto, Rita Santamaria, Filiberto Cimino, Francesco Salvatore

Istituto Nazionale Tumori Fondazione G. Pascale

Research output: Contribution to journal › Article › peer-review

Abstract

The tRNA modifying enzyme, S-adenosylmethionine:tRNA(guanine-7-)-methyltransferase, has been extensively purified from Salmonella typhimurium. A rapid and efficient purification method using phosphocellulose chromatography followed by ammonium sulfate precipitation and Sephadex G-100 gel filtration is described. The enzyme appears to be a single polypeptide chain with a molecular weight of approximately 25 000-30 000 daltons. The K_m for S-adenosylmethionine and for undermethylated tRNA is 53 μ M and 3.4 μM, respectively. The methylation reaction is dependent on added monovalent or divalent cations; 5 mM spermidine, 3 mM MgCl₂ and 1 mM spermine are the most effective. The enzyme, though not homogeneous, is free from contaminating ribonucleases and other tRNA methyltransferases.

Original language	English
Pages (from-to)	97-106
Number of pages	10
Journal	Molecular and Cellular Biochemistry
Volume	52
Issue number	2
DOIs	https://doi.org/10.1007/BF00224919
Publication status	Published - Sep 1983

ASJC Scopus subject areas

Molecular Biology
Genetics
Clinical Biochemistry
Cell Biology

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10.1007/BF00224919

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title = "Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium",

abstract = "The tRNA modifying enzyme, S-adenosylmethionine:tRNA(guanine-7-)-methyltransferase, has been extensively purified from Salmonella typhimurium. A rapid and efficient purification method using phosphocellulose chromatography followed by ammonium sulfate precipitation and Sephadex G-100 gel filtration is described. The enzyme appears to be a single polypeptide chain with a molecular weight of approximately 25 000-30 000 daltons. The Km for S-adenosylmethionine and for undermethylated tRNA is 53 μ M and 3.4 μM, respectively. The methylation reaction is dependent on added monovalent or divalent cations; 5 mM spermidine, 3 mM MgCl2 and 1 mM spermine are the most effective. The enzyme, though not homogeneous, is free from contaminating ribonucleases and other tRNA methyltransferases.",

author = "Alfredo Colonna and Gennaro Ciliberto and Rita Santamaria and Filiberto Cimino and Francesco Salvatore",

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T1 - Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium

AU - Colonna, Alfredo

AU - Ciliberto, Gennaro

AU - Santamaria, Rita

AU - Cimino, Filiberto

AU - Salvatore, Francesco

PY - 1983/9

Y1 - 1983/9

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Isolation and characterization of a tRNA(guanine-7-)-methyltransferase from Salmonella typhimurium

Abstract

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