Sequence‐specific NMR assignments of the trp repressor from Escherichia coli using three‐dimensional 15N/1H heteronuclear techniques

Katherine L.B. BORDEN, Christopher J. BAUER, Thomas A. FRENKIEL, Pamela BECKMANN, Andrew N. LANE

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Sequence‐specific 15N and 1H assignments for the trp holorepressor from Escherichia coli are reported. The trp repressor consists of two identical 107‐residue subunits which are highly helical in the crystal state [Schevitz, R., Otwinowski, Z., Joachimiak, A., Lawson, C. L. & Sigler, P. B. (1985) Nature 317, 782–786]. The high helical content and the relatively large size of the protein (Mr= 25000) make it difficult to assign even the main‐chain resonances by conventional homonuclear two‐dimensional NMR methods. However, we have now assigned the main‐chain resonances of 94% of the residues by using three‐dimensional 15N/1H heteronuclear experiments on a sample of protein uniformly labelled with 15N. The additional resolution obtained by spreading out the signals into three dimensions proved indispensable in making these assignments. In particular, we have been able to resolve signals from residues in the N‐terminal region of the A helix for the first time in solution. The observed NOE results confirm that the repressor is highly helical in solution, and contains no extended chain conformations.

Original languageEnglish
Pages (from-to)137-146
Number of pages10
JournalEuropean Journal of Biochemistry
Volume204
Issue number1
DOIs
StatePublished - Feb 1992

ASJC Scopus subject areas

  • Biochemistry

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