Selective super(15)N labeling of barstar in a T7 polymerase system

The T7 RNA polymerase system for selective protein labeling, previously optimized for the time of introducing the super(15)N amino acid relative to the inducer (IPTG) and host polymerase inhibitor (rifampicin), was further improved by the use of a special growth medium enriched in particular amino a...

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Bibliographic Details
Published inBiophysics (Oxford) Vol. 52; no. 1; pp. 13 - 15
Main Authors Lopukhov, LV, Ponomareva, A A, Yagodina, LO
Format Journal Article
LanguageEnglish
Published 01.02.2007
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Summary:The T7 RNA polymerase system for selective protein labeling, previously optimized for the time of introducing the super(15)N amino acid relative to the inducer (IPTG) and host polymerase inhibitor (rifampicin), was further improved by the use of a special growth medium enriched in particular amino acids and by administration of an aminotransferase inhibitor (a-aminooxyacetic acid); the latter prevented isotope redistribution (as required by NMR studies) even with readily metabolized leucine. The approach is shown to be successful for selective labeling of barstar with [ super(15)N]Trp and [ super(15)N]Leu; it can be used with any proteins expressed in the T7 system.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
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ISSN:0006-3509
1555-6654
DOI:10.1134/S0006350907010034