Then, many clinical investigations and laboratory observations indicated that cortisol, rather cortisone, is the active glucocorticoid,

and 11_-hydroxyl group of the steroid is essential for its anti-inflammatory activity (Bush et al., 1968a; Sarett, 1959). It transpired that cortisone requires the activation in vivo to exert its anti-inflammatory action, and this led to the discovery of 11_-hydroxysteroid dehydrogenase (11_HSD) (Monder and Shackleton, 1984). The metabolic activation of the 11-keto group of cortisone into the 11_-hydroxyl group of cortisol and the reverse reaction to inactivate cortisol is catalyzed by 11_-HSD. The enzyme in many tissues including liver and lung functions in both directions, while in some tissues such as placenta it functions as only a dehydrogenase (Monder and Shackleton, 1984). This paradox was solved after two isoforms of 11_-HSD were cloned. 11_-HSD isoform 1 (11_-HSD1) was first cloned from rat liver in 1989 (Agarwal et al., 1989), and isoform 2 (11_-HSD2) was cloned from sheep and human kidney five years later (Agarwal et al., 1994; Albiston et al., 1994). 11_-HSD1 (Agarwal et al., 1989) and 11_-HSD2 (Agarwal et al., 1994; Albiston et al., 1994) are both microsomal enzymes. However, they possess distinct biochemical properties. 11_-HSD1 is a low-affinity (with Km 2 _M for cortisol) and high capacity enzyme (Monder and White, 1993a). 11_-HSD1 catalyzes both direction, and uses NADP+ as cofactor for oxidase direction and NADPH as cofactor for reductase direction. 11_-HSD1 is present in many tissues including liver, lung, kidney, testis, brain, fat, and the immune system (Monder and White, 1993b). However, 11_- HSD2 is a high affinity enzyme (Km 25 nM for cortisol) (White et al., 1997a). 11_-HSD2 catalyzes unidirectional oxidation of cortisol, and uses NAD+ as cofactor. 11_-HSD2 is primarily present in mineralocorticoid targeted tissue to protect the mineralocorticoid receptor from the excessive binding by cortisol, by deactivating it to cortisone. This protection mechanism and the possible relationship with apparent mineralocorticoid excess syndrome due to mutations in the gene for this enzyme has been well reviewed by White et al. (1997b). More and more evidence points to the fact that the enzymes 11_-HSD1 and 11_-HSD2 also can perform multiple functions, involving in the metabolism of toxins and drugs. 11_-HSD2 is more specific for endogenous glucocorticoids, however, a study showed that 11_-HSD2 catalyzed the metabolism of dexamethasone (Best et al., 1997). 11_-HSD1 has been found to be not only for the metabolism of 11_-hydroxy or 11-keto steroids, but also for the metabolism of 7_-hydroxy, 7_-hydroxy and 7-keto steroids. A comprehensive understanding about this enzyme has been well reviewed by Tomlinson et al. (2004). The present study mainly focuses on the nature of 11_-HSD1 and its involvement of metabolism of steroids, toxins and drugs. 2. 11_-hydroxysteroid dehydrogenase 1 2.1. The structure of 11-HSD1 11_-HSD1 is a member of alcohol short-chain dehydrogenases/reductase (SDR) superfamily, a family with an Nterminal nucleotide cofactor-binding domain (Rossmann-fold) and a catalytic active site in the central region (Oppermann et al., 1997). The nucleotide cofactor-binding region of the family contains GXXXGXG motif that binds specifically to NADP+, NAD+, NADPH, or NADH (Oppermann et al., 1997). The catalytic active site of the family members contains tyrosine (Y or Tyr) and lysine (K or Lys) residues that bind to their ligands (Oppermann et al., 1997). There exists a great sequence similarity of 11_-HSD1 among species (Fig. 1). Typically, the primary structure of 11_-HSD1 of various species contains nucleotide cofactor-binding motif (GASKGIG) and the catalytic active site (YSASK), which is conserved in the SDR family. Furthermore, it has been suggested that Tyr177 of human 11_-HSD1 does not donate a hydrogen bond in substrate binding and determine substrate specificity, but takes part in hydrophobic interaction (Kim et al., 2006). Besides these motifs, the 11_-HSD1 peptide contains a single hydrophobic N-terminal extension which possibly anchors in the smooth endoplasmic reticulum (SER). Using attached FLAG epitopes at the N- and C-terminals of human 11_-HSD1 to determine the enzyme topology, Odermatt et al. (1999) demonstrated that the majority of the peptide resides in the luminal space of the SER with only five amino acids on the cytosolic side and the catalytic domain-containing C terminus protrudes into the SER lumen. The amino acid Lys5 but not Lys6 of 11_HSD1 peptide is critical for the topology of 11_-HSD1. Mutation of Lys5 to Ser5 inverted the orientation of 11_-HSD1 in the SER (Odermatt et al., 1999). Glu25 and Glu26 in 11_-HSD1 are also responsible for determining its luminal orientation. The location of these residues rather than net charge distribution on either side of the helix is critical for membrane topology of 11_-HSD1 (Frick et al., 2004). However, the catalytic activity is not altered after mutation (Odermatt et al., 1999). N-terminally truncated form of rat 11_-HSD1 loses its catalytic activity (Mercer et al., 1993; Obeid and White, 1992), indicating that extensive transmembrane domain might also affect 3D structure for the catalytic active site of the enzyme. 11_-HSD1 can form important disulfide bonds within the 11_-HSD1 second peptide structure and the SER luminal milieu promotes the formation of disulfide bonds (Ozols, 1995b). Purified rat liver 11_-HSD1 is found to be a glycosylated protein (Lakshmi and Monder, 1988). After rat 11_HSD1 gene (Hsd11b1) was cloned and expressed in cultured cells using recombinant vaccinia virus, partial inhibition of glycosylation decreased its oxidase activity by 50% without affecting reductase activity (Agarwal et al., 1990). Two potential N-linked glycosylation sites, Asn158X-Ser160 and Asn203-X-Ser205, were identified in the translated peptide (Agarwal et al., 1990). The role of the two potential glycosylation sites was further investigated in Chinese hamster ovary