Sterolsulphate sulphohydrolase from human placenta microsomes--30 kDa molecular weight form of cholesterol sulphate sulphohydrolase.
The cholesterol sulphate sulphohydrolase (CHS-ase) exhibiting molecular weight of 30 kDa was purified from human placenta microsomes. The microsomal proteins were extracted with 0.5% Triton X-100. The DEAE-cellulose chromatography of the solubilized microsomal proteins, performed at pH 7.6 allowed to separate two enzymatically active fractions. One of them was associated with the protein fraction unbound by DEAE-cellulose, the other was tightly bound by ion exchanger. The 30 kDa cholesterol sulphate sulphohydrolase was purified to homogenity from the protein fraction tightly bound by DEAE-cellulose. The highly purified enzyme preparation (specific activity 385 nmol min(-1)mg(-1) of protein) exhibited optimal activity at pH 6.4, the K(m) was established to be 6.7 x 10(-6)M, the pI value was 7.4. The 30 kDa cholesterol sulphate sulphohydrolase, in contrast to the CHS-ase form originated from the protein fraction unbound by DEAE-cellulose, was not sensitive to alkaline phosphatase treatment and phosphohydrolase inhibitors. The effects of steroids, -SH reacting agents and sulphohydrolase inhibitors on the enzyme activity were tested.
Norkowska
Marzenna
M
Biochemistry Department, Institute of Biology and Molecular Biology, Mikołaj Kopernik University, Gagarina, Poland.
Gniot-Szulzycka
Jadwiga
J
D000469:Alkaline Phosphatase / Q000494:pharmacology; D001192:Arylsulfatases / Q000737:chemistry* / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D017830:Octoxynol / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010920:Placenta / Q000201:enzymology* / Q000378:metabolism; D043266:Steryl-Sulfatase
D004791:Enzyme Inhibitors; D017830:Octoxynol; D000469:Alkaline Phosphatase; D010744:Phosphoric Monoester Hydrolases; C466373:cholesterol sulfate sulfohydrolase; D001192:Arylsulfatases; D043266:Steryl-Sulfatase