Expression of the human genes for steroid 21-hydroxylase and its C169R mutant in insect cells and functional analysis of the expression products |
| |
Authors: | Yu. V. Grischuk P. M. Rubtsov S. N. Beljelarskaya |
| |
Affiliation: | (1) Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia |
| |
Abstract: | Steroid 21-hydroxylase (CYP21A2) is a key enzyme of glucocorticoid and mineralocorticoid biosynthesis in the adrenal cortex and belongs to the family of microsomal cytochrome P450. CYP21A2 deficiency is the most common cause of human congenital adrenal hyperplasia (CAH). Human CYP21A2 and its C169R mutant, observed in a patient with classic CAH, were expressed in Sf9 and Hi5 insect cells infected with recombinant baculoviruses. Functional CYP21A2 was produced to 28% of the total microsomal protein under optimal conditions. The C169R mutation did not affect the efficiency of CYP21A2 synthesis in insect cells, nor did it prevent CYP21A2 incorporation in membranes of the endoplasmic reticulum. Functional analysis in vitro showed that the mutant enzyme almost completely lacked the catalytic activity towards two substrates, progesterone and 17-hydroxyprogesterone. |
| |
Keywords: | steroid 21-hydroxylase congenital adrenal hyperplasia missense mutation recombinant baculovirus insect cells |
本文献已被 SpringerLink 等数据库收录! |