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The Phe300Gln mutant is not activated by bicarbonate concentrations <2 mM. Above 2 mM bicarbonate, ACCO activity increases in a linear (non-saturable) manner, indicating that the binding site for bicarbonate is not functional. Phe300 cannot bind bicarbonate. The Phe300Gln Asc Ka = 0.52 mM and is a high-affinity binding site for Asc, indicating that Phe300 is essential for bicarbonate activation of ACCO.
