Cloning and expression in vitro of human xanthine dehydrogenase/oxidase

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Abstract
To study the expression of human xanthine dehydrogenase/ oxidase (hXDH/XO), we cloned the cDNA covering its complete coding sequence and characterized it by translation in vitro in rabbit reticulocyte lysates and by transient expression in COS-1 cells. Two specific protein products with approximate molecular masses of 150 and 130 kDa were detected in both expression systems. These products are compatible with the molecular sizes of XDH/XO, and these peptides also showed immunoreactivity with polyclonal anti-hXDH antibodies. Significant XDH/XO enzyme activity (277±54 pmol/min per mg of protein) was measured in lysates of transfected COS cells, whereas in control transfections the activities were below the detection limit of our assay (0.2 pmol/min per mg of protein). The COS cells expressed the enzyme predominantly (89.8±0.3%) in the dehydrogenase form.