Essential arginine residues in maize starch synthase IIa are involved in both ADP-glucose and primer binding

Abstract

The arginine-specific reagent phenylglyoxal inactivated the activity of maize starch synthase IIa (SSIIa), due to the modification of at least one arginine residue out of a possible 42. The addition of ADPGlc completely protected SSIIa from the inactivation, indicating that arginine may be involved in the interaction of this anionic substrate with SSIIa. However, site-directed mutagenesis of the conserved Arg-214 in SSIIa showed that this amino acid is important for apparent affinity of SSIIa for its primer (amylopectin and glycogen), as evidenced by a marked increase in the K m for primer upon substitution of this amino acid with no concomitant change in V max, K m for ADPGlc, or secondary structure. Therefore, Arg-214 of SSIIa appears to play a role in its primer binding.