Selenomethionine labeling of recombinant proteins

Methods Mol Biol. 2007:389:165-74. doi: 10.1007/978-1-59745-456-8_12.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids
  • Crystallization
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescence
  • Gene Expression
  • Genetic Vectors
  • Pichia
  • Recombinant Proteins / analysis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Selenomethionine / metabolism*
  • Staining and Labeling*
  • Static Electricity

Substances

  • Amino Acids
  • Recombinant Proteins
  • Selenomethionine