Biochemical and biophysical characterisation yields insights into the mechanism of a Cd/Zn transporting ATPase purified from the hyperaccumulator plant Thlaspi caerulescens

► TcHMA4, a Cd/Zn-ATPase has been purified from roots in its native state. ► TcHMA4 displayed highest activity with submicromolar concentrations of Cd2+, Cu2+ and Zn2+. ► Concentration-dependent changes in activation energy were different for the three metals tested. ► The temperature optimum for ATPase activity was reached at approximately 40 °C for all tested metals. ► EXAFS measurements showed that Cd in TcHMA4 is bound by S from cysteines and not by N from histidines.