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- W41902871 abstract "Previous research on nickel-induced teratogenesis in Xenopus laevis identified several Ni{sup 2+}-binding proteins, including pNiXa in Xenopus ovaries, unfertilized eggs, and embryos. A major goal of this research project is elucidating the role of pNiXa in the uptake, embryotoxicity, and teratogenicity of Ni{sup 2+} in Xenopus. To purify and identify pNiXa, ovarian tissue from mature Xenopus females was homogenized in 3 vol of Tris buffer and centrifuged. The supernatant was centrifuged; the ultracentrifugal supernatant was batch-adsorbed onto DEAE-cellulose. The pNiSa remained unbound and was subsequently adsorbed on phosphocellulose and eluted by a step-wise NaCl gradient. The pNiXa was eluted in 0.25 M NaCl; this fraction was concentrated, and further purified by reverse phase chromatography on a 5 {mu}m C-8 column, with a linear trifluoroacetic acid/acetonitrile gradient. The pNiXa was eluted at {approximately}56% acetonitrile, yielding a single protein band with mol wt {approximately}47 kD,based on SDS-PAGE analysis. Comparison of the amino acid composition of pNiXa versus the results obtained by automated Edman degradation indicated that the N-terminus of pNiXa was blocked. Sequencing of peptide fragments of pNiXa is underway." @default.
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- W41902871 date "1991-03-11" @default.
- W41902871 modified "2023-09-28" @default.
- W41902871 title "Purification of a Ni sup 2+ -binding protein, pNiXa, from Xenopus ovary" @default.
- W41902871 hasPublicationYear "1991" @default.
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