Molecular cloning and functional expression of the cytochrome P450 11B-hydroxylase of the guinea pig

Hannes E. Bülow; Katharina Möbius; Volker Bähr; Rita Bernhardt

doi:10.1006/bbrc.1996.0591

Molecular cloning and functional expression of the cytochrome P450 11B-hydroxylase of the guinea pig

Hannes E. Bülow, Katharina Möbius, Volker Bähr, Rita Bernhardt

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Abstract

A cDNA library has been constructed from adrenal glands of guinea pigs receiving a low sodium/high potassium diet. From this library a complementary DNA encoding an 11β-hydroxylase (EC 1.14.15.4) was isolated. Northern blot analysis revealed a major transcript of 2.2 kb size. The cDNA was cloned into an eucaryotic expression vector and the activity of the enzyme towards various substrates was investigated after expression in COS-1 cells. The enzyme was able to perform 11β-hydroxylations of 11-deoxycorticosterone, 11-deoxycortisol and androstendione as well as 18-hydroxylations and 19-hydroxylations to a small extent. Phylogenetic analyses with thirteen protein sequences of the CYP11B subfamily showed an early divergence of the guinea pig, but could not establish a definite branching order.

Original language	English (US)
Pages (from-to)	304-312
Number of pages	9
Journal	Biochemical and Biophysical Research Communications
Volume	221
Issue number	2
DOIs	https://doi.org/10.1006/bbrc.1996.0591
State	Published - Apr 16 1996
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Molecular cloning and functional expression of the cytochrome P450 11B-hydroxylase of the guinea pig",

abstract = "A cDNA library has been constructed from adrenal glands of guinea pigs receiving a low sodium/high potassium diet. From this library a complementary DNA encoding an 11β-hydroxylase (EC 1.14.15.4) was isolated. Northern blot analysis revealed a major transcript of 2.2 kb size. The cDNA was cloned into an eucaryotic expression vector and the activity of the enzyme towards various substrates was investigated after expression in COS-1 cells. The enzyme was able to perform 11β-hydroxylations of 11-deoxycorticosterone, 11-deoxycortisol and androstendione as well as 18-hydroxylations and 19-hydroxylations to a small extent. Phylogenetic analyses with thirteen protein sequences of the CYP11B subfamily showed an early divergence of the guinea pig, but could not establish a definite branching order.",

author = "B{\"u}low, {Hannes E.} and Katharina M{\"o}bius and Volker B{\"a}hr and Rita Bernhardt",

note = "Funding Information: We thank Dr. K. Denner and B. B{\"o}ttner for helpful discussions and valuable advice in HPLC and transfection experiments. We also express our gratitude to D. Fiedler for expert sequencing. This work was supported by the Deutsche Forschungsgemeinschaft Grant DFG Be 13436-1.",

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doi = "10.1006/bbrc.1996.0591",

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AU - Bülow, Hannes E.

AU - Möbius, Katharina

AU - Bähr, Volker

AU - Bernhardt, Rita

N1 - Funding Information: We thank Dr. K. Denner and B. Böttner for helpful discussions and valuable advice in HPLC and transfection experiments. We also express our gratitude to D. Fiedler for expert sequencing. This work was supported by the Deutsche Forschungsgemeinschaft Grant DFG Be 13436-1.

PY - 1996/4/16

Y1 - 1996/4/16

N2 - A cDNA library has been constructed from adrenal glands of guinea pigs receiving a low sodium/high potassium diet. From this library a complementary DNA encoding an 11β-hydroxylase (EC 1.14.15.4) was isolated. Northern blot analysis revealed a major transcript of 2.2 kb size. The cDNA was cloned into an eucaryotic expression vector and the activity of the enzyme towards various substrates was investigated after expression in COS-1 cells. The enzyme was able to perform 11β-hydroxylations of 11-deoxycorticosterone, 11-deoxycortisol and androstendione as well as 18-hydroxylations and 19-hydroxylations to a small extent. Phylogenetic analyses with thirteen protein sequences of the CYP11B subfamily showed an early divergence of the guinea pig, but could not establish a definite branching order.

AB - A cDNA library has been constructed from adrenal glands of guinea pigs receiving a low sodium/high potassium diet. From this library a complementary DNA encoding an 11β-hydroxylase (EC 1.14.15.4) was isolated. Northern blot analysis revealed a major transcript of 2.2 kb size. The cDNA was cloned into an eucaryotic expression vector and the activity of the enzyme towards various substrates was investigated after expression in COS-1 cells. The enzyme was able to perform 11β-hydroxylations of 11-deoxycorticosterone, 11-deoxycortisol and androstendione as well as 18-hydroxylations and 19-hydroxylations to a small extent. Phylogenetic analyses with thirteen protein sequences of the CYP11B subfamily showed an early divergence of the guinea pig, but could not establish a definite branching order.

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Molecular cloning and functional expression of the cytochrome P450 11B-hydroxylase of the guinea pig

Abstract

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