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

  • Koenig B
  • Rogowski M
  • Louis J
  • 31

    Readers

    Mendeley users who have this article in their library.
  • 35

    Citations

    Citations of this article.

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 alpha-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.

Author-supplied keywords

  • E. coli
  • Fusion protein
  • Isotope labeling
  • NMR
  • Recombinant peptide

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Bernd W. Koenig

  • Marco Rogowski

  • John M. Louis

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free