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Selenomethionine Labeling of Recombinant Proteins
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.
Affiliation(s): (2) Department of Cell and Molecular Biology, University of Uppsala, Uppsala, Sweden
Book Title: Pichia Protocols
Series: Methods in Molecular Biology  |  Volume: 389  |  Pub. Date: Aug-08-2007  |  Page Range: 165-174  |  DOI: 10.1007/978-1-59745-456-8_12
Subject:  Biotechnology
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