title: Selective production of rat mutant selenoprotein W with and without bound glutathione.
abstract: Matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) and electrospray ionization mass spectrometry (ESI MS) analysis of a 6x His-tagged recombinant form of rat mutant selenoprotein W (RMSW) reveals that aerobic growth conditions primarily produce a form of RMSW without bound glutathione (10,305 Da) whereas anaerobic conditions produce a glutathione-bound (305 Da) form (10,610 Da). Purification of RMSW was achieved with a procedure employing acetone precipitation and DEAE-cellulose chromatography, in addition to Ni-NTA agarose chromatography. Additional steps, including polyvalent metal ion binding (PMIB) resin chromatography and CM-cellulose chromatography, were necessary after elution from the Ni-NTA agarose column, in order to maintain solubility of the purified protein.
authors.affiliation: Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR 97331, USA.
authors.forename: A T
authors.initials: AT
authors.lastname: Bauman
authors.forename: D A
authors.initials: DA
authors.lastname: Malencik
authors.forename: D F
authors.initials: DF
authors.lastname: Barofsky
authors.forename: E
authors.initials: E
authors.lastname: Barofsky
authors.forename: S R
authors.initials: SR
authors.lastname: Anderson
authors.forename: P D
authors.initials: PD
authors.lastname: Whanger
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D009154:Mutation; D011485:Protein Binding; D014176:Protein Biosynthesis; D020033:Protein Isoforms; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D051151:Selenoprotein W; D051140:Selenoproteins; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D020033:Protein Isoforms; D011506:Proteins; D011994:Recombinant Proteins; D051151:Selenoprotein W; D051140:Selenoproteins; D005978:Glutathione