Cholesterol sulphate sulphohydrolase from human placenta microsomes--purification and properties of the dephosphorylated form of enzyme

The procedure for purification of cholesterol sulphate sulphohydrolase (ChS-ase) from human placenta microsomes was elaborated. The highy purified enzyme preparation (specific activity 2000 nmol×min^?1×mg protein^?1) exhibited optimal activity at pH 9.0. The K_m value was established to be 1.5±0.85×10^?5 M. The high molecular weight form (200 kDa) and the low molecular weight form (20 kDa) of the enzyme were separated. The interconversion of the high molecular weight variant into the low one occurs under the influence of dephosphorylation. Both forms exhibited typical Michaelis–Menten saturation kinetics. The effect of different compounds on the enzyme activity was tested.