This Article Full Text Full Text (PDF) Submit a related Letter to the Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Grischuk, Y. Articles by Krone, N. Search for Related Content PubMed PubMed Citation Articles by Grischuk, Y. Articles by Krone, N. Pubmed/NCBI databases Gene GEO Profiles HomoloGene UniGene Substance via MeSH Genetics Home Reference Related Collections Adrenal and Hypertension

Four Novel Missense Mutations in the CYP21A2 Gene Detected in Russian Patients Suffering from the Classical Form of Congenital Adrenal Hyperplasia: Identification, Functional Characterization, and Structural Analysis

Engelhardt Institute of Molecular Biology (Y.G., P.R., S.B., V.Pr.), Russian Academy of Sciences, Moscow 119991, Russian Federation; Division of Pediatric Endocrinology, Department of Pediatrics (F.G.R., W.G.S., N.K.), Christian-Albrechts-Universität zu Kiel, Universitätsklinikum Schleswig-Holstein (Campus Kiel), D-24105 Kiel, Germany; Biochemisches Institut (J.G.), Christian-Albrechts-Universität zu Kiel, D-24098 Kiel, Germany; and Institute of Pediatric Endocrinology, Endocrinological Research Center (N.K., T.S., V.Pe., A.T.), Moscow 117036, Russian Federation

Address all correspondence and requests for reprints to: Nils Krone, M.D., Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics, Christian-Albrechts-Universität zu Kiel, Universitätsklinikum Schleswig-Holstein (Campus Kiel), Schwanenweg 20, D-24105 Kiel, Germany. E-mail: krone{at}pediatrics.uni-kiel.de.

Context: Congenital adrenal hyperplasia is a group of autosomal recessive inherited disorders of steroidogenesis. The most frequent cause is the deficiency of steroid 21-hydroxylase (CYP21) due to mutations in the CYP21A2 gene.

Objective: We analyzed the functional and structural consequences of the four CYP21A2 missense mutations (C169R, G178R, W302R, and R426C) to prove their clinical relevance and study their impact on CYP21 function.

Results: Analyzing the mutations in vitro revealed an almost absent or negligible CYP21 activity for the conversion of 17-hydroxyprogesterone to 11-deoxycortisol and progesterone to deoxycorticosterone. Protein translation and intracellular localization were not affected by the mutants, as could be demonstrated by Western blotting and immunofluorescence studies. Analysis of these mutants in a three-dimensional model structure of the CYP21 protein explained the observed in vitro effects because all the mutations severely interfere either directly or indirectly with important structures of the 21-hydroxylase protein.

Conclusion: The in vitro expression analysis of residual enzyme function is a complementary method to genotyping and an important tool for improving the understanding of the clinical phenotype of 21-hydroxylase deficiency. This forms the foundation for accurate clinical and genetic counseling and for prenatal diagnosis and treatment. Moreover, this report demonstrates that the combination of in vitro enzyme analysis and molecular modeling can yield novel insights into CYP450 structure-functional relationships.

This article has been cited by other articles:

				V. Dhir, N. Reisch, C. M. Bleicken, J. Lebl, C. Kamrath, H.-P. Schwarz, J. Grotzinger, W. G. Sippell, F. G. Riepe, W. Arlt, et al. Steroid 17{alpha}-Hydroxylase Deficiency: Functional Characterization of Four Mutations (A174E, V178D, R440C, L465P) in the CYP17A1 Gene J. Clin. Endocrinol. Metab., August 1, 2009; 94(8): 3058 - 3064. [Abstract] [Full Text] [PDF]

				M. Welzel, N. Wustemann, G. Simic-Schleicher, H. G. Dorr, E. Schulze, G. Shaikh, P. Clayton, J. Grotzinger, P.-M. Holterhus, and F. G. Riepe Carboxyl-Terminal Mutations in 3{beta}-Hydroxysteroid Dehydrogenase Type II Cause Severe Salt-Wasting Congenital Adrenal Hyperplasia J. Clin. Endocrinol. Metab., April 1, 2008; 93(4): 1418 - 1425. [Abstract] [Full Text] [PDF]