Selective backbone labeling of proteins using 1,2-13C2-pyruvate as carbon source

J Biomol NMR. 2009 Jul;44(3):167-73. doi: 10.1007/s10858-009-9326-y. Epub 2009 May 26.

Abstract

A simple isotope labeling approach for selective (13)C/(15)N backbone labeling of proteins is described. Using {1,2-(13)C(2)}-pyruvate as the sole carbon source in bacterial growth media, selective incorporation of (13)C(alpha)-(13)CO spin-pairs into the backbones of protein molecules with medium-to-high levels of (13)C-enrichment is possible for a subset of 12 amino acids. The isotope labeling scheme has been tested on a pair of proteins--a 7-kDa immunoglobulin binding domain B1 of streptococcal protein G and an 82-kDa enzyme malate synthase G. A number of protein NMR applications are expected to benefit from the {1,2-(13)C(2)}-pyruvate based protein production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Carbon Isotopes / chemistry*
  • Magnetic Resonance Spectroscopy / methods*
  • Malate Synthase / chemistry
  • Models, Biological
  • Proteins / chemistry*
  • Pyruvic Acid / chemistry*

Substances

  • Bacterial Proteins
  • Carbon Isotopes
  • IgG Fc-binding protein, Streptococcus
  • Proteins
  • Pyruvic Acid
  • Malate Synthase