Selenomethionine labeling of recombinant proteins.
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.
Larsson
Anna M
AM
Department of Cell and Molecular Biology, University of Uppsala, Uppsala, Sweden.
Jones
T Alwyn
TA
D000596:Amino Acids; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D015870:Gene Expression; D005822:Genetic Vectors; D010843:Pichia; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D012645:Selenomethionine / Q000378:metabolism*; D013194:Staining and Labeling; D055672:Static Electricity
D000596:Amino Acids; D011994:Recombinant Proteins; D012645:Selenomethionine