Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance

In order to study the intracellular polyamine distribution in Escherichia coli, 13C-NMR spectra of [1,4-13C]putrescine were obtained after addition of the latter to intact bacteria. The 13C-enriched methylene signal underwent line broadening. When the cells were centrifuged after 90 min the cell-bou...

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Autores principales:	Frydman, Benjamín Jaime, Goldemberg, Sara Hebe, Algranati, Israel David
Publicado:	1984
Materias:	(E. coli) 13C-NMR Putrescine distribution bacterial DNA carbon putrescine transfer RNA article Escherichia coli methodology nuclear magnetic resonance spectroscopy ribosome Carbon Isotopes DNA, Bacterial Magnetic Resonance Spectroscopy Putrescine Ribosomes RNA, Transfer
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674889_v805_n4_p337_Frydman http://hdl.handle.net/20.500.12110/paper_01674889_v805_n4_p337_Frydman
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_01674889_v805_n4_p337_Frydman
record_format	dspace
spelling	paper:paper_01674889_v805_n4_p337_Frydman2023-06-08T15:16:31Z Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance Frydman, Benjamín Jaime Goldemberg, Sara Hebe Algranati, Israel David (E. coli) 13C-NMR Putrescine distribution bacterial DNA carbon putrescine transfer RNA article Escherichia coli methodology nuclear magnetic resonance spectroscopy ribosome Carbon Isotopes DNA, Bacterial Escherichia coli Magnetic Resonance Spectroscopy Putrescine Ribosomes RNA, Transfer In order to study the intracellular polyamine distribution in Escherichia coli, 13C-NMR spectra of [1,4-13C]putrescine were obtained after addition of the latter to intact bacteria. The 13C-enriched methylene signal underwent line broadening. When the cells were centrifuged after 90 min the cell-bound putrescine peak had a linewidth of 23 Hz, while the supernatant liquid showed an unbound putrescine signal with a linewidth smaller than 1 Hz. By using 13C-enriched internal standards it could be shown that the linewidening was not due to the heterogeneity of the medium or to an in vivo paramagnetic effect. Cell-bound putrescine was liberated by addition of trichloroacetic acid and was therefore non-covalently linked to macromolecular cell structures. Cell-bound [13C]putrescine could be displaced by addition of an excess of [12C]putrescine. When samples of membranes, soluble protein, DNA, tRNA and ribosomes from E. coli were incubated with [1,4-13C]putrescine, strong binding was detected only in the ribosomal and membrane fractions. The ribosome-putrescine complex showed properties similar to those determined with the intact cells. By measuring the nuclear Overhauser enhancements η, it was possible to estimate that only about 50% of the polyamine was linked to the macromolecules. Determination of the T1 values of free and ribosomal-bound putrescine allowed the calculation of a correlation time, τc = 4·10-7 s for the latter. T1 and τc value for the ribosome-putrescine complex were those expected for a motional regime of slowly tumbling molecules. © 1984. Fil:Frydman, B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Goldemberg, S.H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Algranati, I.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1984 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674889_v805_n4_p337_Frydman http://hdl.handle.net/20.500.12110/paper_01674889_v805_n4_p337_Frydman
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	(E. coli) 13C-NMR Putrescine distribution bacterial DNA carbon putrescine transfer RNA article Escherichia coli methodology nuclear magnetic resonance spectroscopy ribosome Carbon Isotopes DNA, Bacterial Escherichia coli Magnetic Resonance Spectroscopy Putrescine Ribosomes RNA, Transfer
spellingShingle	(E. coli) 13C-NMR Putrescine distribution bacterial DNA carbon putrescine transfer RNA article Escherichia coli methodology nuclear magnetic resonance spectroscopy ribosome Carbon Isotopes DNA, Bacterial Escherichia coli Magnetic Resonance Spectroscopy Putrescine Ribosomes RNA, Transfer Frydman, Benjamín Jaime Goldemberg, Sara Hebe Algranati, Israel David Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance
topic_facet	(E. coli) 13C-NMR Putrescine distribution bacterial DNA carbon putrescine transfer RNA article Escherichia coli methodology nuclear magnetic resonance spectroscopy ribosome Carbon Isotopes DNA, Bacterial Escherichia coli Magnetic Resonance Spectroscopy Putrescine Ribosomes RNA, Transfer
description	In order to study the intracellular polyamine distribution in Escherichia coli, 13C-NMR spectra of [1,4-13C]putrescine were obtained after addition of the latter to intact bacteria. The 13C-enriched methylene signal underwent line broadening. When the cells were centrifuged after 90 min the cell-bound putrescine peak had a linewidth of 23 Hz, while the supernatant liquid showed an unbound putrescine signal with a linewidth smaller than 1 Hz. By using 13C-enriched internal standards it could be shown that the linewidening was not due to the heterogeneity of the medium or to an in vivo paramagnetic effect. Cell-bound putrescine was liberated by addition of trichloroacetic acid and was therefore non-covalently linked to macromolecular cell structures. Cell-bound [13C]putrescine could be displaced by addition of an excess of [12C]putrescine. When samples of membranes, soluble protein, DNA, tRNA and ribosomes from E. coli were incubated with [1,4-13C]putrescine, strong binding was detected only in the ribosomal and membrane fractions. The ribosome-putrescine complex showed properties similar to those determined with the intact cells. By measuring the nuclear Overhauser enhancements η, it was possible to estimate that only about 50% of the polyamine was linked to the macromolecules. Determination of the T1 values of free and ribosomal-bound putrescine allowed the calculation of a correlation time, τc = 4·10-7 s for the latter. T1 and τc value for the ribosome-putrescine complex were those expected for a motional regime of slowly tumbling molecules. © 1984.
author	Frydman, Benjamín Jaime Goldemberg, Sara Hebe Algranati, Israel David
author_facet	Frydman, Benjamín Jaime Goldemberg, Sara Hebe Algranati, Israel David
author_sort	Frydman, Benjamín Jaime
title	Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance
title_short	Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance
title_full	Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance
title_fullStr	Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance
title_full_unstemmed	Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance
title_sort	putrescine distribution in escherichia coli studied in vivo by 13c nuclear magnetic resonance
publishDate	1984
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674889_v805_n4_p337_Frydman http://hdl.handle.net/20.500.12110/paper_01674889_v805_n4_p337_Frydman
work_keys_str_mv	AT frydmanbenjaminjaime putrescinedistributioninescherichiacolistudiedinvivoby13cnuclearmagneticresonance AT goldembergsarahebe putrescinedistributioninescherichiacolistudiedinvivoby13cnuclearmagneticresonance AT algranatiisraeldavid putrescinedistributioninescherichiacolistudiedinvivoby13cnuclearmagneticresonance
_version_	1768543556043014144

Putrescine distribution in Escherichia coli studied in vivo by 13C nuclear magnetic resonance

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