Quemener E, Amet Y, Fournier G, Di Stefano S, Abalain J H, Floch H H
Département de Biochimie et Biologie Moléculaire, Faculté de Médecine, Brest, France.
Biochem Biophys Res Commun. 1994 Nov 30;205(1):269-74. doi: 10.1006/bbrc.1994.2660.
5 alpha-reductase 2 from human prostate solubilized into an active and stable form using a non-ionic detergent octyl glucoside was successfully purified using a four-step chromatographic procedure. The enzyme was obtained as an apparently homogeneous protein exhibiting an apparent molecular weight of 42 kDa upon SDS-PAGE. Con A, DBA, UEA-I, and RCA60 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 enzyme had no effect. These results indicate that the human prostate testosterone 5 alpha-reductase 2 is an O-glycosylated sialoglycoprotein with a peptide moiety of about 30 kDa; the oligosaccharide side chains contain mannose, N-acetyl galactosamine, fucose, galactose and sialic acids.
使用非离子去污剂辛基葡糖苷将人前列腺中的5α-还原酶2溶解成活性和稳定形式，通过四步色谱法成功纯化。该酶以一种明显均一的蛋白质形式获得，在SDS-PAGE上显示表观分子量为42 kDa。伴刀豆球蛋白A、双花扁豆凝集素、荆豆凝集素I和蓖麻凝集素60识别该蛋白质。用O-糖苷酶和神经氨酸酶处理后，出现了一种表观分子量约为30 kDa的蛋白质。另一方面，用N-糖苷酶处理该酶没有效果。这些结果表明，人前列腺睾酮5α-还原酶2是一种O-糖基化的唾液糖蛋白，其肽部分约为30 kDa；寡糖侧链含有甘露糖、N-乙酰半乳糖胺、岩藻糖、半乳糖和唾液酸。
