Human l-3-phosphoserine phosphatase: sequence, expression and evidence for a phosphoenzyme intermediate

AbstractWe report the sequence of the cDNA encoding human l-3-phosphoserine phosphatase. The encoded polypeptide contains 225 residues and shows 30% sequence identity with the Escherichia coli enzyme. The human protein was expressed in a bacterial expression system and purified. Similar to known l-3-phosphoserine phosphatases, it catalyzed the Mg2+-dependent hydrolysis of l-phosphoserine and an exchange reaction between l-serine and l-phosphoserine. In addition we found that the enzyme was phosphorylated upon incubation with l-[32P]phosphoserine, which indicates that the reaction mechanism proceeds via the formation of a phosphoryl-enzyme intermediate. The sensitivity of the phosphoryl-enzyme to alkali and to hydroxylamine suggests that an aspartyl- or a glutamyl-phosphate was formed. The nucleotide sequence of the cDNA described in this article has been deposited in the EMBL data base under accession number Y10275