Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C4/C18 column system

Jeroen R. Mesters; Erik L.H. Vorstenbosch; Aldo J. de Boer; Barend Kraal

doi:10.1016/0021-9673(94)80314-5

Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C₄/C₁₈ column system

Jeroen R. Mesters, Erik L.H. Vorstenbosch, Aldo J. de Boer, Barend Kraal^*

^*Corresponding author for this work

Institute of Biochemistry

Leiden University

3 Citations (Scopus)

Abstract

Phe-tRNA^Phe, N-acetyl-Phe-tRNA^Phe and Leu-tRNA^Leu-4 (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C₄ column and inverse salt gradient elution. The alterations at the 3′ end of the tRNAs result in a considerable shift of retention time on this column. The method is useful not only for obtaining tRNA preparations as required for poly(U) translational studies, but also for producing 20-50-mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated species (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.

Original language	English
Journal	Journal of Chromatography A
Volume	679
Issue number	1
Pages (from-to)	93-98
Number of pages	6
ISSN	0021-9673
DOIs	https://doi.org/10.1016/0021-9673(94)80314-5
Publication status	Published - 09.09.1994

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/0021-9673(94)80314-5

Cite this

@article{38c0a2b637cc42fc88001ff46f39e55d,

title = "Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C4/C18 column system",

abstract = "Phe-tRNAPhe, N-acetyl-Phe-tRNAPhe and Leu-tRNALeu-4 (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C4 column and inverse salt gradient elution. The alterations at the 3′ end of the tRNAs result in a considerable shift of retention time on this column. The method is useful not only for obtaining tRNA preparations as required for poly(U) translational studies, but also for producing 20-50-mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated species (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.",

author = "Mesters, {Jeroen R.} and Vorstenbosch, {Erik L.H.} and {de Boer}, {Aldo J.} and Barend Kraal",

year = "1994",

month = sep,

day = "9",

doi = "10.1016/0021-9673(94)80314-5",

language = "English",

volume = "679",

pages = "93--98",

journal = "Journal of Chromatography A",

issn = "0021-9673",

number = "1",

}

Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C₄/C₁₈ column system. / Mesters, Jeroen R.; Vorstenbosch, Erik L.H.; de Boer, Aldo J. et al.
In: Journal of Chromatography A, Vol. 679, No. 1, 09.09.1994, p. 93-98.

Research output: Journal Articles › Journal articles › Research › peer-review

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T1 - Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C4/C18 column system

AU - Mesters, Jeroen R.

AU - Vorstenbosch, Erik L.H.

AU - de Boer, Aldo J.

AU - Kraal, Barend

PY - 1994/9/9

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N2 - Phe-tRNAPhe, N-acetyl-Phe-tRNAPhe and Leu-tRNALeu-4 (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C4 column and inverse salt gradient elution. The alterations at the 3′ end of the tRNAs result in a considerable shift of retention time on this column. The method is useful not only for obtaining tRNA preparations as required for poly(U) translational studies, but also for producing 20-50-mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated species (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.

AB - Phe-tRNAPhe, N-acetyl-Phe-tRNAPhe and Leu-tRNALeu-4 (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C4 column and inverse salt gradient elution. The alterations at the 3′ end of the tRNAs result in a considerable shift of retention time on this column. The method is useful not only for obtaining tRNA preparations as required for poly(U) translational studies, but also for producing 20-50-mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated species (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.

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