Purification and properties of arylsulphatase A from rabbit testis

Abstract

Rabbit testis arylsulphatase A was purified 140-fold with a recovery of20 % from detergent extracts of an acetone-dried powder by using DE-52 cellulose column chromatography, gel filtration on Sephadex G-200 and preparative isoelectric focusing. The purified enzyme showed one major band with one minor contaminant on electrophoresis in a 7.5% (w/v) polyacrylamide gel at pH 8.3. On sodium dodecyl sulphate/polyacrylamidegel electrophoresis, a single major band was observed with minor contaminants. The final preparation of enzyme was free from general proteolytic, esterase, hyaluronidase, A6-glucuronidase and fi-galactosidase activities. Rabbit testicular arylsulphatase A exists as a dimer of mol.wt. 110000 at pH7

Isolation and characterization of the pig endometrial arylsulphatase A.

“…The duration of the incubation and the incubation temperature influence the kinetic parameters of arylsulphatase A (Luijten et al, 1978), and this makes exact comparison of these values with the literature values somewhat difficult. However, the values obtained in the present work are in the range of those obtained with other arylsulphatases A (Yang & Srivastava, 1976;Farooqui & Srivastava, 1979).…”

“…The isoelectric point of the ox liver enzyme is pH 3.4 (Nichol & Roy, 1964). Most other arylsulphatases A, including those from human urine (Stevens et al, 1973), liver (Neuwelt et al, 1972) and rabbit testis (Yang & Srivastava, 1976), show a pl value of 4.6, which is very near to the pI value of the endometrial enzyme. The relatively high content of potential Vol.…”

“…The optima were at pH 5.5 and 5.1, which are in the usual range of pH optima for arylsulphatases A from other sources (Yang & Srivastava, 1976).…”

“…For activity determinations unstained gels were cut into 25 mm segments and incubated in the assay system for arylsulphatase for 30-120 min. Determination of relative molecular mass The Mr of the enzyme was determined by gel filtration on a Sephadex G-200 column (48 cm x 2cm) as described by Yang & Srivastava (1976). The enzyme was assumed to be a globular protein for derivation of the Mr.…”

“…The isoelectric focusing was performed on an LKB electrofocusing column as described by Yang & Srivastava (1976). Isoelectric focusing was performed at 50C at 2-3W for 48 h with a starting voltage of 500V.…”