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^*

^*Korrespondierende/r Autor/-in für diese Arbeit

Institut für Biochemie

Leiden University

6 Zitate (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.

Originalsprache	Englisch
Zeitschrift	Journal of Chromatography A
Jahrgang	679
Ausgabenummer	1
Seiten (von - bis)	93-98
Seitenumfang	6
ISSN	0021-9673
DOIs	https://doi.org/10.1016/0021-9673(94)80314-5
Publikationsstatus	Veröffentlicht - 09.09.1994

Fördermittel

E.L.H.V. was supported by a grant from the Netherlands Foundation for Chemical research (SON).

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Forschungsschwerpunkt: Infektion und Entzündung - Zentrum für Infektions- und Entzündungsforschung Lübeck (ZIEL)

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10.1016/0021-9673(94)80314-5

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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,

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

language = "English",

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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, Jahrgang 679, Nr. 1, 09.09.1994, S. 93-98.

Publikation: Beiträge in Fachzeitschriften › Zeitschriftenaufsätze › Forschung › Begutachtung

TY - JOUR

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

Y1 - 1994/9/9

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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DO - 10.1016/0021-9673(94)80314-5

M3 - Journal articles

AN - SCOPUS:0028085789

SN - 0021-9673

VL - 679

SP - 93

EP - 98

JO - Journal of Chromatography A

JF - Journal of Chromatography A

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