Purification of the testicular galactolipid: 3'-phosphoadenosine 5'-phosphosulfate sulfotransferase.
The rat testicular galactolipid sulfotransferase has been purified by affinity chromatography using 3'5'-adenosine diphosphate-agarose affinity chromatography. Both galactosyl glycerolipid and galactosyl ceramide were effective substrates with Km values of 4.8 and 1.1 microM respectively. A single protein of molecular mass 56 kDa was present in the purified sulfotransferase preparation as monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. Specific photoaffinity substrate labeling, using an azido derivative of galactosyl ceramide, was used to identify this protein, both in crude extracts and when purified. The protein was also selectively phosphorylated in the presence of the rat testicular galactolipid sulfotransferase stimulatory protein kinase.
Sakac
D
D
Research Institute, Hospital for Sick Children, Ontario, Canada.
Zachos
M
M
Lingwood
C A
CA
D000818:Animals; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D017830:Octoxynol; D010766:Phosphorylation; D011092:Polyethylene Glycols; D051381:Rats; D011919:Rats, Inbred Strains; D015238:Sulfotransferases / Q000302:isolation & purification* / Q000378:metabolism; D013737:Testis / Q000201:enzymology*
D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol; C022321:PAPS sulfotransferase; D015238:Sulfotransferases; C049108:galactolipid sulfotransferase