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Published on: November 3, 2018
Cloning, expression, purification, and some properties of calf purine nucleoside phosphorylase
Katarzyna Stepniak1, Agnieszka Girstun, Beata Wielgus-Kutrowska
1Department of Biophysics, Institute of Experimental Physics, Warsaw University, Warsaw, Poland.
Abstract:
Calf spleen purine nucleoside phosphorylase (PNP) is considered a model enzyme for the trimeric PNPs subfamily. PCR amplification of the calf phosphorylase from the calf spleen library, cloning, overexpression of the recombinant PNP, its enzymatic activity and interactions with typical ligands of mammalian wild type PNP are described. Relative activity of the recombinant phosphorylase versus several substrates is similar to the respective values obtained for the enzyme isolated from calf spleen. As for the nonrecombinant calf PNP, the unusual fluorescence properties of the PNP/guanine complex were observed and characterized.
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