Isoleucine 259 and isoleucine 260 residues in Streptococcus gordonii soluble inorganic pyrophosphatase play an important role in enzyme activity

The hinge region of the family II soluble inorganic pyrophosphatase (PPase) found in Streptococcus gordonii DL1 (Challis) has previously been shown to play an important role in opening and closing of the active site between the N- and C-terminal domains of PPase. Amino acid residues isoleucine 259 (I259) and isoleucine 260 (I260) are highly conserved among catalytically active family II PPases and are located very close to the hinge region. Substitution of either I259 or I260 with a hydrophilic acidic amino acid (glutamate or aspartate) resulted in adverse effects on the kinetic properties of the enzyme. The I259/E and I259/D variants were nearly catalytically inactive (k(cat)/K-m