GLYCOSYLATED NATURE OF TESTOSTERONE 5-ALPHA-REDUCTASE 2 PURIFIED FROMHUMAN PROSTATE

5 alpha-reductase 2 from human prostate solubilized into an active and stable form using a non-ionic detergent octylglucoside was successful ly purified using a four-step chromatographic procedure. The enzyme wa s obtained as an apparently homogeneous protein exhibiting an apparent molecular weight of 42 kDa upon SDS-PAGE. Con A, DBA, UEA-I, and RCA6 0 lectins recognized this protein. After treatment with O-glycosidase and neuraminidase, a protein of an apparent molecular weight about 30 kDa appeared. On the other hand, N-glycosidase treatment of this enzym e had no effect. These results indicate that the human prostate testos terone Sa-reductase 2 is an O-glycosylated sialoglycoprotein with a pe ptide moiety of about 30 kDa; the oligosaccharide side chains contain mannose, N-acetyl galactosamine, fucose, galactose and sialic acids. ( C) 1994 Academic Press, Inc.