The better tag remains unseen

J Am Chem Soc. 2008 Nov 12;130(45):14932-3. doi: 10.1021/ja806212j. Epub 2008 Oct 21.

Abstract

One of the most crucial steps in protein structure determination by nuclear magnetic resonance (NMR) spectroscopy is the preparation of highly concentrated and well behaving protein samples. Here we present a system of modular tags which allows for high level expression, sophisticated purification of full-length protein, and solubility enhancement while keeping the amount of additional resonances low. This system consists of two different expression constructs and utilizes the tight binding of human calmodulin (hCaM) to the calmodulin binding peptide (CBP), which has already been used as a purification tag.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Calmodulin / chemistry*
  • Calmodulin-Binding Proteins / chemistry*
  • Glutathione Transferase / chemistry
  • Humans
  • Mice
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Peptide Fragments / chemistry
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification

Substances

  • Calmodulin
  • Calmodulin-Binding Proteins
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • Glutathione Transferase