Selenomethionine Labeling of Recombinant Proteins

  • Anna M. Larsson
  • T. Alwyn Jones
Part of the Methods in Molecular Biology book series (MIMB, volume 389)

Abstract

Selenomethionine incorporation is a standard method for determining the phases in protein crystallography by single- or multiwavelength anomalous dispersion. Recombinant expression of selenomethionine-containing protein in non-auxotrophic Pichia pastoris strains yield an incorporation of about 50%. The expression of a mutated variant of Penicillium minioluteum dextranase in P. pastoris is used to illustrate the method utilized to obtain selenomethionyl-substituted protein and to show the phasing power of the acquired anomalous signal. The dextranase structure was solved using the anomalous signal achieved from 50% selenomethionine incorporation.

Key Words

Dextranase MAD multiwavelength anomalous dispersion Pichia pastoris SeMet selenomethionine 

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Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Anna M. Larsson
    • 1
  • T. Alwyn Jones
    • 1
  1. 1.Department of Cell and Molecular BiologyUniversity of UppsalaUppsalaSweden

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