Comparison of different transition metal ions for immobilized metal affinity chromatography of selenoprotein P from human plasma.
Cu2+, Ni2+, Zn2+, Co2+ and Cd2+ were evaluated in metal ion affinity chromatography for enrichment of selenoprotein P, and immobilized Co2+ affinity chromatography was found to be the most selective chromatographic method. The chromatography was performed by fast protein liquid chromatography and the fractionation was followed by analysis of the collected fractions for selenium by inductively coupled plasma mass spectrometry. By the combination of immobilized Co2+ affinity chromatography and heparin affinity chromatography a simple method was developed yielding a 14,800-fold enrichment of selenoprotein P. The purity of the protein was determined by SDS-PAGE and by sequencing from polyvinylidene difluoride blots of SDS-PAGE gels.
Sidenius
U
U
Department of Analytical and Pharmaceutical Chemistry, The Royal Danish School of Phannacy, Copenhagen O. ulsi@mail.dfh.dk
Farver
O
O
Jøns
O
O
Gammelgaard
B
B
D000345:Affinity Labels; D000595:Amino Acid Sequence; D002104:Cadmium; D002846:Chromatography, Affinity / Q000379:methods*; D003035:Cobalt; D003300:Copper; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008670:Metals; D008970:Molecular Weight; D009532:Nickel; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D051149:Selenoprotein P; D051140:Selenoproteins; D012685:Sepharose / Q000031:analogs & derivatives; D015032:Zinc
D000345:Affinity Labels; D008670:Metals; D011506:Proteins; D051149:Selenoprotein P; D051140:Selenoproteins; C037405:heparin-sepharose; D002104:Cadmium; D003035:Cobalt; D003300:Copper; D009532:Nickel; D012685:Sepharose; D015032:Zinc