title: Electroanalytical determination of tungsten and molybdenum in proteins.
abstract: Recent crystal structure determinations accelerated the progress in the biochemistry of tungsten-containing enzymes. In order to characterize these enzymes, a sensitive determination of this metal in protein-containing samples is necessary. An electroanalytical tungsten determination has successfully been adapted to determine the tungsten and molybdenum content in enzymes. The tungsten and molybdenum content can be measured simultaneously from 1 to 10 microg of purified protein with little or no sample handling. More crude protein samples require precipitation of interfering surface active material with 10% perchloric acid. This method affords the isolation of novel molybdenum- and tungsten-containing proteins via molybdenum and tungsten monitoring of column fractions, without using radioactive isotopes. A screening of soluble proteins from Pyrococcus furiosus for tungsten, using anion-exchange column chromatography to separate the proteins, has been performed. The three known tungsten-containing enzymes from P. furiosus were recovered with this screening.
authors.affiliation: Kluyver Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands. p.l.hagedoorn@tnw.tudelft.nl
authors.forename: P L
authors.initials: PL
authors.lastname: Hagedoorn
authors.forename: P
authors.initials: P
authors.lastname: van't Slot
authors.forename: H P
authors.initials: HP
authors.lastname: van Leeuwen
authors.forename: W R
authors.initials: WR
authors.lastname: Hagen
mesh_terms: D011232:Chemical Precipitation; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004563:Electrochemistry / Q000379:methods*; D004798:Enzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008667:Metalloproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008982:Molybdenum / Q000032:analysis*; D010472:Perchlorates / Q000378:metabolism; D020027:Pyrococcus furiosus / Q000201:enzymology; D012995:Solubility; D014414:Tungsten / Q000032:analysis*
chemical_list: D004798:Enzymes; D008667:Metalloproteins; D010472:Perchlorates; D008982:Molybdenum; D014414:Tungsten