15N-labeled P22 c2 repressor for nuclear magnetic resonance studies of protein-DNA interactions |
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Authors: | H Senn A Eugster G Otting F Suter K Wüthrich |
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Institution: | (1) Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule, ETH-Hönggerberg, CH-8093 Zürich, Switzerland;(2) Present address: Pharmaceutical Division, Preclinical Research, Sandoz Ltd., CH-4002 Basel, Switzerland |
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Abstract: | The salmonella phage P22 c2 repressor was produced with 90% 15N isotope labeling of all leucines, using the expression system E. coli W3110 lac I
Q/pTP 125. The N-terminal DNA-binding domain 1–76 was obtained by chymotrypsin cleavage. Its characterization by biochemical techniques, mass spectrometry, and one- and two-dimensional nuclear magnetic resonance (NMR) showed that highly residue-selective isotope labeling was achieved with the minimal growth medium used. The ability to obtain such isotope labeling opens new avenues for NMR studies of protein-DNA interactions in the P22 operator system. |
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Keywords: | Isotope labeling nuclear magnetic resonance protein conformation protein-DNA interactions P22c2 repressor |
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