Comparative analysis of two glyceraldehyde-3-phosphate dehydrogenases from a thermoacidophilic archaeon, Sulfolobus tokodaii

► We purified glyceraldehyde-3-phosphate: NADP+ oxidoreductase from Sulfolobus tokodaii. ► St-GAPN showed K-type allosteric activation by glucose-1-phosphate. ► We identified the residues that determine the co-substrate preference of St-GAPN. ► St-GAPDH showed high gluconeogenic activity with unique co-substrate usage.