Modulation of testicular galactolipid sulphotransferase activity by phosphorylation. Stimulation of enzyme activity in vitro by an endogenous kinase.
Evidence is presented for a testicular protein kinase activity capable of stimulating the activity in vitro of a partially purified preparation of the testicular galactolipid sulphotransferase. This enzyme is responsible for the synthesis of the major mammalian testicular glycolipid, sulphogalactosylglycerol, and is an early marker of differentiation during spermatogenesis. This stimulatory activity has been separated by affinity chromatography, using 3',5'-ADP-agarose, from both the detergent-solubilized microsomes (microsomal fractions) and the soluble fraction of the testicular homogenate. The stimulator was eluted from the affinity matrix by either a salt, or, more selectively, a cyclic AMP gradient. Thus this matrix can function as an analogue of 3',5'-cyclic AMP. The activity of the sulphotransferase stimulator was ATP-dependent and coincident with protein kinase activity. Sulphotransferase activity was also stimulated in the presence of commercial preparations of cyclic AMP-dependent protein kinase and stimulation was prevented in the presence of kinase inhibitors. Our results suggest that sulphogalactolipid biosynthesis is regulated by a phosphorylation process during spermatogenesis. In addition, our results suggest that affinity chromatography on 3',5'-ADP-agarose may provide a general method for the purification of cyclic AMP-dependent kinases.
Sakac
D
D
Research Institute, Hospital for Sick Children, Toronto, Ont., Canada.
Lingwood
C A
CA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002846:Chromatography, Affinity; D004789:Enzyme Activation; D008297:Male; D010766:Phosphorylation; D011494:Protein Kinases / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000378:metabolism*; D013737:Testis / Q000187:drug effects / Q000201:enzymology*
D000255:Adenosine Triphosphate; D011494:Protein Kinases; D015238:Sulfotransferases; C049108:galactolipid sulfotransferase