What common structural features and variations of mammalian P450s are known to date?

Otyepka, Michal; Skopalík, Josef; Anzenbacherová, Eva; Anzenbacher, Pavel

doi:10.1016/j.bbagen.2006.09.013

Cited by 123 publications

(120 citation statements)

References 77 publications

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“…Helix C is most obviously involved in redox-partner binding, and the disturbance of this contact can cause slower substrate conversion due to insufficient electron flux. 19,20 The siblings 2 and 3 originate from northern Italy. Both were heterozygous for the mutation p.(Leu299Pro) and the novel p.(Arg332Trp) mutant.…”

Section: Discussionmentioning

confidence: 99%

Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: functional consequences of four CYP11B1 mutations

et al. 2013

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Congenital adrenal hyperplasia (CAH) is one of the most common autosomal recessive inherited endocrine disease. Steroid 11b-hydroxylase deficiency (11b-OHD) is the second most common form of CAH. The aim of the study was to study the functional consequences of three novel and one previously described CYP11B1 gene mutations (p.(Arg143Trp), p.(Ala306Val), p.(Glu310Lys) and p.(Arg332Gln)) detected in patients suffering from classical and non-classical 11b-OHD. Functional analyses were performed by using a HEK293 cell in vitro expression system comparing wild type (WT) with mutant 11b-hydroxylase activity. Mutant proteins were examined in silico to study their effect on the three-dimensional structure of the protein. Two mutations (p.(Ala306Val) and p.(Glu310Lys)) detected in patients with classical 11b-OHD showed a nearly complete loss of 11b-hydroxylase activity. The mutations p.(Arg143Trp) and p.(Arg332Gln) detected in patients with non-classical 11b-OHD showed a partial functional impairment with approximately 8% and 6% of WT activity, respectively. Functional mutation analysis allows the classification of novel CYP11B1 mutations as causes of classical and non-classical 11b-OHD. The detection of patients with non-classical phenotypes underscores the importance to screen patients with a phenotype comparable to non-classical 21-hydroxylase deficiency for mutations in the CYP11B1 gene in case of a negative analysis of the CYP21A2 gene. As CYP11B1 mutations are most often individual for a family, the in vitro analysis of novel mutations is essential for clinical and genetic counselling.

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Section: Discussionmentioning

confidence: 99%

Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: functional consequences of four CYP11B1 mutations

et al. 2013

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“…CYPs101 [4], 119 [5], 158A2 [6], 2B4 [7], 2C5 [8], and 2C9 [9]). Most pronounced motions are usually observed in the substrate binding regions, including B/C and F/G segments, so that the shape of the substrate binding cavity becomes adjustable to the shape of the corresponding substrate [2,[10][11][12].…”

mentioning

confidence: 99%

Conformational dynamics in the F/G segment of CYP51 from Mycobacterium tuberculosis monitored by FRET

Lepesheva

Seliškar

Knutson

et al. 2007

Archives of Biochemistry and Biophysics

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A cysteine was introduced into the FG-loop (P187C) of CYP51 from Mycobacterium tuberculosis (MT) for selective labeling with BODIPY and fluorescence energy transfer (FRET) analysis. Forster radius for the BODIPY-heme pair was calculated assuming that the distance between the heme and Cys187 in solution corresponds to that in the crystal structure of ligand free MTCYP51. Interaction of MTCYP51 with azole inhibitors ketoconazole and fluconazole or the substrate analog estriol did not influence the fluorescence, but titration with the substrate lanosterol quenched BODIPY emission, the effect being proportional to the portion of substrate-bound MTCYP51. The detected changes correspond to ~10 Å decrease in the calculated distance between BODIPY-Cys187 and the heme. The results confirm 1) functional importance of conformational motions in the MTCYP51 F/G segment and 2) applicability of FRET to monitor them in solution. KeywordsCytochrome P450; sterol 14α-demethylase; ligand binding; conformational changes; FRET The cytochrome P450 superfamily currently includes more than 6,300 enzymes (http://drnelson.utmem.edu/CytochromeP450.html) oxidizing a wide variety of xenobiotics (compounds from the environment) and endogenous substrates. Even at less than 16% amino acid identity, all P450s preserve a common overall structural fold (α-helical with orthogonal bundle architecture according to CATH classification [1]) and the set of secondary structural elements that allow the superfamily to metabolize a wide variety of diverse and unrelated structures. By now it has become quite generally accepted that this enormous catalytic versatility arises from the P450 conformational flexibility [2,3]. Ligandinduced conformational changes encompassing small to large movements are seen in the Xray structures of several bacterial and drug-metabolizing mammalian CYPs 1 (e.g. CYPs101 * Corresponding author: Michael R. Waterman, Tel.: 615-343-1373; Fax: 615-322-4349; E-mail: michael.waterman@vanderbilt.edu.Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. . On the other hand, site-directed mutagenesis of five amino acid residues in this region (L172, G175, P178, R194 and D195), which are highly conserved in the CYP51 family and exposed into the substrate binding cavity of MTCYP51 has shown that all of them are essential for MTCYP51 catalytic activity at the stage of the interaction with substrate. Moreover, mutation of A197 in the middle of the MTCYP51 G-helix to the helix-breaking glycine causes about one order of magnitude enzyme activation and sharp increase in the substrate bound (high-spin) po...

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“…The frameshift mutation p.His125Thrfs*8 terminates the translation at the B-C loop of the protein (30). By this, all relevant enzyme structures including the substrate recognitions sides and haeme-binding sites are not present, resulting in a complete loss of function (31,32).…”

Section: Molecular Modellingmentioning

confidence: 99%

“…Two leucines are inserted in a leucine-rich part of the L-helix carrying four leucines. The L-helix is involved in haeme binding as well as in interactions with redox partners (30). The distal L-helix forms parts of the protein's surface.…”

Section: Molecular Modellingmentioning

confidence: 99%

Characterisation of three novel CYP11B1 mutations in classic and non-classic 11β-hydroxylase deficiency

Polat

Kulle

Karaca

et al. 2014

European Journal of Endocrinology

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Background: Congenital adrenal hyperplasia (CAH) is one of the most common autosomal recessive inherited endocrine diseases. Steroid 11b-hydroxylase (P450c11) deficiency (11OHD) is the second most common form of CAH. Aim: The aim of the study was to study the functional consequences of three novel CYP11B1 gene mutations (p.His125Thrfs*8, p.Leu463_Leu464dup and p.Ser150Leu) detected in patients suffering from 11OHD and to correlate this data with the clinical phenotype. Methods: Functional analyses were done by using a HEK293 cell in vitro expression system comparing WT with mutant P450c11 activity. Mutant proteins were examined in silico to study their effect on the three-dimensional structure of the protein.Results: Two mutations (p.His125Thrfs*8 and p.Leu463_Leu464dup) detected in patients with classic 11OHD showed a complete loss of P450c11 activity. The mutation (p.Ser150Leu) detected in a patient with non-classic 11OHD showed partial functional impairment with 19% of WT activity. Conclusion: Functional mutation analysis enables the correlation of novel CYP11B1 mutations to the classic and non-classic 11OHD phenotype respectively. Mutations causing a non-classic phenotype show typically partial impairment due to reduced maximum reaction velocity comparable with non-classic mutations in 21-hydroxylase deficiency. The increasing number of mutations associated with non-classic 11OHD illustrate that this disease should be considered as diagnosis in patients with otherwise unexplained hyperandrogenism.

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What common structural features and variations of mammalian P450s are known to date?

Cited by 123 publications

References 77 publications

Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: functional consequences of four CYP11B1 mutations

Congenital adrenal hyperplasia due to 11-beta-hydroxylase deficiency: functional consequences of four CYP11B1 mutations

Conformational dynamics in the F/G segment of CYP51 from Mycobacterium tuberculosis monitored by FRET

Characterisation of three novel CYP11B1 mutations in classic and non-classic 11β-hydroxylase deficiency

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