| An efficient and cost-effective isotope labeling protocol for proteins expressed in Escherichia coli. | |
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MedLine Citation:
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PMID: 9566315 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A cost-effective protocol for uniform 15N and/or 13C isotope labeling of bacterially expressed proteins is presented. Unlike most standard protocols, cells are initially grown in a medium containing nutrients at natural abundance and isotopically labeled nutrients are only supplied at the later stages of growth and during protein expression. This permits the accumulation of a large cell mass without the need to employ expensive isotopically labeled nutrients. The abrupt decrease in oxygen consumption that occurs upon complete exhaustion of essential nutrients is used to precisely time the switch between unlabeled and labeled nutrients. Application of the protocol is demonstrated for wild-type and a mutant of the N-terminal zinc-binding domain of HIV-1 integrase. |
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Authors:
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M Cai; Y Huang; K Sakaguchi; G M Clore; A M Gronenborn; R Craigie |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Journal of biomolecular NMR Volume: 11 ISSN: 0925-2738 ISO Abbreviation: J. Biomol. NMR Publication Date: 1998 Jan |
Date Detail:
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Created Date: 1998-06-01 Completed Date: 1998-06-01 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9110829 Medline TA: J Biomol NMR Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 97-102 Citation Subset: IM; X |
Affiliation:
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Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Carbon Isotopes Cost-Benefit Analysis Escherichia coli / genetics HIV Integrase / biosynthesis, chemistry, genetics Magnetic Resonance Spectroscopy Nitrogen Isotopes Point Mutation Recombinant Proteins / biosynthesis, chemistry*, genetics |
| Chemical | |
Reg. No./Substance:
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0/Carbon Isotopes; 0/Nitrogen Isotopes; 0/Recombinant Proteins; EC 2.7.7.-/HIV Integrase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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