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Cell-free synthesis of 15 N-labeled proteins for NMR studies

Ozawa, Kiyoshi; Dixon, Nicholas; Otting, Gottfried

Description

Modern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15N-labeled proteins from 15N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR spectra recorded of the crude reaction mixture without concentration or chromatographic purification of the protein. We review methodological aspects of cell-free in...[Show more]

dc.contributor.authorOzawa, Kiyoshi
dc.contributor.authorDixon, Nicholas
dc.contributor.authorOtting, Gottfried
dc.date.accessioned2015-12-13T22:45:42Z
dc.identifier.issn1521-6543
dc.identifier.urihttp://hdl.handle.net/1885/79911
dc.description.abstractModern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15N-labeled proteins from 15N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR spectra recorded of the crude reaction mixture without concentration or chromatographic purification of the protein. We review methodological aspects of cell-free in vitro protein synthesis based on an Escherichia coli cell extract, in particular with regard to the production of 15N-labeled proteins for analysis by NMR spectroscopy.
dc.publisherTaylor & Francis Group
dc.sourceIUBMB Life
dc.subjectKeywords: amino acid; cell extract; nitrogen 15; protein; amino acid metabolism; amino terminal sequence; cell free system; chromatography; cost; economic aspect; equipment; Escherichia coli; genetic heterogeneity; isotope labeling; methodology; nonhuman; nuclear m 15N-HSQC spectrum; 15N-labeling; Cell-free protein synthesis; E. coli S30 extract; NMR spectroscopy
dc.titleCell-free synthesis of 15 N-labeled proteins for NMR studies
dc.typeJournal article
local.description.notesImported from ARIES
local.description.refereedYes
local.identifier.citationvolume57
dc.date.issued2005
local.identifier.absfor100299 - Environmental Biotechnology not elsewhere classified
local.identifier.ariespublicationMigratedxPub8272
local.type.statusPublished Version
local.contributor.affiliationOzawa, Kiyoshi, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationDixon, Nicholas, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationOtting, Gottfried, College of Physical and Mathematical Sciences, ANU
local.description.embargo2037-12-31
local.bibliographicCitation.issue9
local.bibliographicCitation.startpage615
local.bibliographicCitation.lastpage622
local.identifier.doi10.1080/15216540500217859
dc.date.updated2015-12-11T10:24:42Z
local.identifier.scopusID2-s2.0-27444447384
CollectionsANU Research Publications

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