Catalytic mechanism and cofactor preference of dihydrodipicolinate reductase from methicillin-resistant Staphylococcus aureus

► This study describes for the first time the catalytic mechanism and cofactor preference of an antibiotic target, MRSA-DHDPR. ► MRSA-DHDPR follows a compulsory order ternary complex kinetic mechanism. ► MRSA-DHDPR binds with 20-fold greater affinity to NADPH compared to NADH. ► However, MRSA-DHDPR shows substrate (DHDP) inhibition when utilizing NADPH as the cofactor. ► These findings provide insight into rational drug design strategies targeting MRSA-DHDPR.