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A rapid method to attain isotope labeled small soluble peptides for NMR studies

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Abstract

A widely applicable strategy is presented for efficient and rapid production of small water soluble peptides expressed as fusion proteins with the immunoglobulin-binding domain of streptococcal protein G. A simple extraction and purification scheme that includes a protease cleavage step to release the target peptide is described. The yield of authentic target peptide exceeds 10 mg per liter of culture. Production of U-13C, 15N and highly deuterated U-13C, 15N isotope labeled peptide is demonstrated for the 11 residue S2 peptide, corresponding to the C-terminus of the α-subunit of transducin, and the coiled coil trimerization domain from cartilage matrix protein (CMPcc), respectively. Heteronuclear two-dimensional NMR spectra are used for initial peptide characterization.

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Authors and Affiliations

  1. Structural Biology Institute, IBI-2, Research Centre Jülich, D-52425, Jülich

    Bernd W. Koenig

  2. Institute of Physical Biology, Heinrich-Heine-Universität, D-40225, Düsseldorf, Germany

    Bernd W. Koenig

  3. Department of Structural Biology, Biozentrum, University of Basel, CH-4056, Basel, Switzerland

    Marco Rogowski

  4. Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Bethesda, MD, 20892-0520, U.S.A

    John M. Louis

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  1. Bernd W. Koenig
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  2. Marco Rogowski
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  3. John M. Louis
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Correspondence to Bernd W. Koenig.

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Koenig, B.W., Rogowski, M. & Louis, J.M. A rapid method to attain isotope labeled small soluble peptides for NMR studies. J Biomol NMR 26, 193–202 (2003). https://doi.org/10.1023/A:1023887412387

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