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Cloning and Characterization of the First Member of the Nudix Family from Arabidopsis thaliana *

The sequence motif commonly called a Nudix box, represented by (GX 5EX 7REVXEEXGU) is the marker of a widely distributed family of enzymes that catalyze the hydrolysis of a variety of nucleoside diphosphate derivatives. Here we describe the cloning and characterization of an Arabidopsis thaliana cDNA encoding a Nudix hydrolase that degrades NADH. The deduced amino acid sequence of AtNUDT1 contains 147 amino acids. The recombinant AtNUDT1 was expressed in Escherichia coliand purified. In the presence of Mn2+ and the optimal pH of 7. 0, the recombinant AtNUDT1 catalyzed the hydrolysis of NADH with aK m value of 0. 36 mm. AV max of 12. 7 units mg −1 for NADH was determined. The recombinant AtNUDT1 migrated as a dimer on a gel filtration column. Biochemical analysis of recombinant AtNUDT1 indicated that the first characterized member of the Nudix family fromA. thaliana is a NADH pyrophosphatase.