Expression of human kidney 11beta-hydroxysteroid dehydrogenase (11-HSD2) in bacteria.
Expression of human kidney 11beta-hydroxysteroid dehydrogenase (11-HSD2) in bacteria.
复制标题
人肾 11β-羟基类固醇脱氢酶 (11-HSD2) 在细菌中的表达。
DOI：
10.1006/bbrc.1999.0259
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
White,PC
中科院分区：
文献类型：
--
作者：
Nunez,BS;Mune,T;White,PC
The kidney isozyme of 11β-hydroxysteroid dehydrogenase (11-HSD2) protects the mineralocorticoid receptor from spurious activation by glucocorticoids. To explore structure-function relationships, human 11-HSD2 cDNA was subcloned into the bacterial expression vector, pET25b.E. colitransformed with wild-type cDNA produced active enzyme that retained biochemical characteristics of the native protein. The addition of 6 histidine residues to the C-terminus of the wild-type enzyme (11-HSD2/His) increased activity 2-fold. Whereas wild-type activity was almost completely sedimented following 100,000gcentrifugation, 10-30% of total activity of 11-HSD2/His remained in the supernatant. The 11-HSD2 isozyme normally contains three N-terminal hydrophobic domains. Mutant 11-HSD2/His possessing a single hydrophobic domain retained partial activity, but elimination of all domains inactivated the enzyme. Thus, the N-terminal hydrophobic domains are essential for complete activity of 11-HSD2 but association with an intact cell membrane is not.
