Molecular cloning and nucleotide sequence of DNA of mitochondrial cytochrome P-450(11β) of bovine adrenal cortex

Ken-Ichirou Morohashi, Hidefumi Yoshioka, Osamu Gotoh, Yoshiie Okada, Kimiko Yamamoto, Toshiyuki Miyata, Kazuhiro Sogawa, Yoshiaki Fuhi-kuriyama, Tsuneo Omura

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

cDNA clones of the mRNA for bovine adrenal cytochrome P-450(11β) were isolated. Sequence analysis of a 4 kb long cDNA revealed the primary structure of P-450(11β), which consisted of 503 amino acids (Mr: 57, 924) and contained an extension peptide of 24 amino acids at the NH2-terminus of the mature P-450(11β) molecule. A bovine genomic DNA containing the 1st exon and its leader sequence of P-450(11β) gene was also isolated from a bovine gene library. Determination of the transcription initiation site by SI nuclease analysis using the cloned genomic DNA confirmed that the methionine codon near the 5' side of the 4 kb long cDNA was the initiation codon. Comparisons of the primary structures among P-45O(11β) and other forms of cytochrome P-450 including P-450(SCC) indicated that the two mitochondrial P-450s, P-450(11β) and P-450(SCC), were significantly different from microsomal forms of cytochrome P-450. The homology between P-45O(11β) and P-450(SCC), was 36%, which is higher than the values between P-450(11β) and various microsoraal P-450s. An alignment of P-45O(11β) and P-450(SCC) to give maximum matching showed four highly conserved regions (C-l, C-2, C-3, and C-4). The homology values of these regions were 58-70%, considerably higher than the overall homology between these two mitochondrial P-450s. A putative heme binding site and a steroid binding site were located in the conserved regions. Hydropathy profiles of P-45O(11β) and P-450(SCC) were very similar. A definite difference was noticed at the NH2terminal portion between mitochondrial and microsomal types of P-450. Microsomal type of cytochrome P-450 had a hydro-phobic sequence consisting of about 20 amino acids, whereas mitochondrial type had an extension peptide containing many positively changed amino acids.

Original languageEnglish
Pages (from-to)559-568
Number of pages10
JournalJournal of biochemistry
Volume102
Issue number3
DOIs
Publication statusPublished - Jan 1 1987

Fingerprint

Cloning
Adrenal Cortex
Molecular Cloning
Mitochondrial DNA
Cytochrome P-450 Enzyme System
Nucleotides
Amino Acids
Complementary DNA
Binding Sites
Peptides
Initiator Codon
Transcription Initiation Site
DNA
Heme
Gene Library
Codon
Methionine
Sequence Analysis
Exons
Clone Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Molecular cloning and nucleotide sequence of DNA of mitochondrial cytochrome P-450(11β) of bovine adrenal cortex. / Morohashi, Ken-Ichirou; Yoshioka, Hidefumi; Gotoh, Osamu; Okada, Yoshiie; Yamamoto, Kimiko; Miyata, Toshiyuki; Sogawa, Kazuhiro; Fuhi-kuriyama, Yoshiaki; Omura, Tsuneo.

In: Journal of biochemistry, Vol. 102, No. 3, 01.01.1987, p. 559-568.

Research output: Contribution to journalArticle

Morohashi, K-I, Yoshioka, H, Gotoh, O, Okada, Y, Yamamoto, K, Miyata, T, Sogawa, K, Fuhi-kuriyama, Y & Omura, T 1987, 'Molecular cloning and nucleotide sequence of DNA of mitochondrial cytochrome P-450(11β) of bovine adrenal cortex', Journal of biochemistry, vol. 102, no. 3, pp. 559-568. https://doi.org/10.1093/oxfordjournals.jbchem.a122089
Morohashi, Ken-Ichirou ; Yoshioka, Hidefumi ; Gotoh, Osamu ; Okada, Yoshiie ; Yamamoto, Kimiko ; Miyata, Toshiyuki ; Sogawa, Kazuhiro ; Fuhi-kuriyama, Yoshiaki ; Omura, Tsuneo. / Molecular cloning and nucleotide sequence of DNA of mitochondrial cytochrome P-450(11β) of bovine adrenal cortex. In: Journal of biochemistry. 1987 ; Vol. 102, No. 3. pp. 559-568.
@article{c8b8e901a2844944b42c949887345be5,
title = "Molecular cloning and nucleotide sequence of DNA of mitochondrial cytochrome P-450(11β) of bovine adrenal cortex",
abstract = "cDNA clones of the mRNA for bovine adrenal cytochrome P-450(11β) were isolated. Sequence analysis of a 4 kb long cDNA revealed the primary structure of P-450(11β), which consisted of 503 amino acids (Mr: 57, 924) and contained an extension peptide of 24 amino acids at the NH2-terminus of the mature P-450(11β) molecule. A bovine genomic DNA containing the 1st exon and its leader sequence of P-450(11β) gene was also isolated from a bovine gene library. Determination of the transcription initiation site by SI nuclease analysis using the cloned genomic DNA confirmed that the methionine codon near the 5' side of the 4 kb long cDNA was the initiation codon. Comparisons of the primary structures among P-45O(11β) and other forms of cytochrome P-450 including P-450(SCC) indicated that the two mitochondrial P-450s, P-450(11β) and P-450(SCC), were significantly different from microsomal forms of cytochrome P-450. The homology between P-45O(11β) and P-450(SCC), was 36{\%}, which is higher than the values between P-450(11β) and various microsoraal P-450s. An alignment of P-45O(11β) and P-450(SCC) to give maximum matching showed four highly conserved regions (C-l, C-2, C-3, and C-4). The homology values of these regions were 58-70{\%}, considerably higher than the overall homology between these two mitochondrial P-450s. A putative heme binding site and a steroid binding site were located in the conserved regions. Hydropathy profiles of P-45O(11β) and P-450(SCC) were very similar. A definite difference was noticed at the NH2terminal portion between mitochondrial and microsomal types of P-450. Microsomal type of cytochrome P-450 had a hydro-phobic sequence consisting of about 20 amino acids, whereas mitochondrial type had an extension peptide containing many positively changed amino acids.",
author = "Ken-Ichirou Morohashi and Hidefumi Yoshioka and Osamu Gotoh and Yoshiie Okada and Kimiko Yamamoto and Toshiyuki Miyata and Kazuhiro Sogawa and Yoshiaki Fuhi-kuriyama and Tsuneo Omura",
year = "1987",
month = "1",
day = "1",
doi = "10.1093/oxfordjournals.jbchem.a122089",
language = "English",
volume = "102",
pages = "559--568",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "3",

}

TY - JOUR

T1 - Molecular cloning and nucleotide sequence of DNA of mitochondrial cytochrome P-450(11β) of bovine adrenal cortex

AU - Morohashi, Ken-Ichirou

AU - Yoshioka, Hidefumi

AU - Gotoh, Osamu

AU - Okada, Yoshiie

AU - Yamamoto, Kimiko

AU - Miyata, Toshiyuki

AU - Sogawa, Kazuhiro

AU - Fuhi-kuriyama, Yoshiaki

AU - Omura, Tsuneo

PY - 1987/1/1

Y1 - 1987/1/1

N2 - cDNA clones of the mRNA for bovine adrenal cytochrome P-450(11β) were isolated. Sequence analysis of a 4 kb long cDNA revealed the primary structure of P-450(11β), which consisted of 503 amino acids (Mr: 57, 924) and contained an extension peptide of 24 amino acids at the NH2-terminus of the mature P-450(11β) molecule. A bovine genomic DNA containing the 1st exon and its leader sequence of P-450(11β) gene was also isolated from a bovine gene library. Determination of the transcription initiation site by SI nuclease analysis using the cloned genomic DNA confirmed that the methionine codon near the 5' side of the 4 kb long cDNA was the initiation codon. Comparisons of the primary structures among P-45O(11β) and other forms of cytochrome P-450 including P-450(SCC) indicated that the two mitochondrial P-450s, P-450(11β) and P-450(SCC), were significantly different from microsomal forms of cytochrome P-450. The homology between P-45O(11β) and P-450(SCC), was 36%, which is higher than the values between P-450(11β) and various microsoraal P-450s. An alignment of P-45O(11β) and P-450(SCC) to give maximum matching showed four highly conserved regions (C-l, C-2, C-3, and C-4). The homology values of these regions were 58-70%, considerably higher than the overall homology between these two mitochondrial P-450s. A putative heme binding site and a steroid binding site were located in the conserved regions. Hydropathy profiles of P-45O(11β) and P-450(SCC) were very similar. A definite difference was noticed at the NH2terminal portion between mitochondrial and microsomal types of P-450. Microsomal type of cytochrome P-450 had a hydro-phobic sequence consisting of about 20 amino acids, whereas mitochondrial type had an extension peptide containing many positively changed amino acids.

AB - cDNA clones of the mRNA for bovine adrenal cytochrome P-450(11β) were isolated. Sequence analysis of a 4 kb long cDNA revealed the primary structure of P-450(11β), which consisted of 503 amino acids (Mr: 57, 924) and contained an extension peptide of 24 amino acids at the NH2-terminus of the mature P-450(11β) molecule. A bovine genomic DNA containing the 1st exon and its leader sequence of P-450(11β) gene was also isolated from a bovine gene library. Determination of the transcription initiation site by SI nuclease analysis using the cloned genomic DNA confirmed that the methionine codon near the 5' side of the 4 kb long cDNA was the initiation codon. Comparisons of the primary structures among P-45O(11β) and other forms of cytochrome P-450 including P-450(SCC) indicated that the two mitochondrial P-450s, P-450(11β) and P-450(SCC), were significantly different from microsomal forms of cytochrome P-450. The homology between P-45O(11β) and P-450(SCC), was 36%, which is higher than the values between P-450(11β) and various microsoraal P-450s. An alignment of P-45O(11β) and P-450(SCC) to give maximum matching showed four highly conserved regions (C-l, C-2, C-3, and C-4). The homology values of these regions were 58-70%, considerably higher than the overall homology between these two mitochondrial P-450s. A putative heme binding site and a steroid binding site were located in the conserved regions. Hydropathy profiles of P-45O(11β) and P-450(SCC) were very similar. A definite difference was noticed at the NH2terminal portion between mitochondrial and microsomal types of P-450. Microsomal type of cytochrome P-450 had a hydro-phobic sequence consisting of about 20 amino acids, whereas mitochondrial type had an extension peptide containing many positively changed amino acids.

UR - http://www.scopus.com/inward/record.url?scp=0023411497&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023411497&partnerID=8YFLogxK

U2 - 10.1093/oxfordjournals.jbchem.a122089

DO - 10.1093/oxfordjournals.jbchem.a122089

M3 - Article

VL - 102

SP - 559

EP - 568

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 3

ER -