Pulmonary cytochrome P450 enzymes belonging to the CYP4B subfamily from an Australian marsupial, the tammar wallaby (Macropus eugenii)

Natalie L. Milic, Suong N.T. Ngo, Ceilidh L. Marchant, Tamara A. Height, Ross A. McKinnon

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Cytochromes P450 (CYPs) are critically important in the oxidative metabolism of a diverse array of xenobiotics and endogenous substrates. We have previously reported the cloning and characterisation of the koala CYP4A15, the first reported member of the CYP4 family from marsupials, and have demonstrated important species differences in CYP4A activity and tissue expression. In the present study, the cloning of CYP4B1 in the wallaby (Macropus eugenii) and their expression across marsupials is described. Rabbit anti-mouse CYP4B1 antibody detected immunoreactive proteins in lung and liver microsomes from all test marsupials, with relative weak signal detected from the koala, suggesting a species-specific expression. Microsomal 2-aminofluorene bio-activation (a CYP4B1 marker) in wallaby lung was comparable to that of rabbit, with significant higher activities detected in wallaby liver and kidneys compared to rabbit. A 1548 bp wallaby lung CYP4B complete cDNA, designated CYP4B1, which encodes a protein of 510 amino acids and shares 72% nucleotide and 69% amino acid sequence identity to human CYP4B1, was cloned by polymerase chain reaction approaches. The results demonstrate the presence of wallaby CYP4B1 that shares several common features with other published CYP4Bs; however the wallaby CYP4B1 cDNA contains four extra amino acid residues at the NH2-terminal, a fundamentally conserved transmembrane anchor of all eukaryote CYPs.

    Original languageEnglish
    Pages (from-to)60-66
    Number of pages7
    JournalComparative Biochemistry and Physiology, Part C
    Volume153
    Issue number1
    DOIs
    Publication statusPublished - 2011

    Fingerprint

    Macropodidae
    Marsupialia
    Cytochrome P-450 Enzyme System
    Lung
    Phascolarctidae
    Cloning
    Rabbits
    Amino Acids
    Liver
    Organism Cloning
    Complementary DNA
    Cytochrome P-450 CYP4A
    Polymerase chain reaction
    Liver Microsomes
    Xenobiotics
    cytochrome P-450 CYP4B1
    Eukaryota
    Anchors
    Metabolism
    Amino Acid Sequence

    Cite this

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    title = "Pulmonary cytochrome P450 enzymes belonging to the CYP4B subfamily from an Australian marsupial, the tammar wallaby (Macropus eugenii)",
    abstract = "Cytochromes P450 (CYPs) are critically important in the oxidative metabolism of a diverse array of xenobiotics and endogenous substrates. We have previously reported the cloning and characterisation of the koala CYP4A15, the first reported member of the CYP4 family from marsupials, and have demonstrated important species differences in CYP4A activity and tissue expression. In the present study, the cloning of CYP4B1 in the wallaby (Macropus eugenii) and their expression across marsupials is described. Rabbit anti-mouse CYP4B1 antibody detected immunoreactive proteins in lung and liver microsomes from all test marsupials, with relative weak signal detected from the koala, suggesting a species-specific expression. Microsomal 2-aminofluorene bio-activation (a CYP4B1 marker) in wallaby lung was comparable to that of rabbit, with significant higher activities detected in wallaby liver and kidneys compared to rabbit. A 1548 bp wallaby lung CYP4B complete cDNA, designated CYP4B1, which encodes a protein of 510 amino acids and shares 72{\%} nucleotide and 69{\%} amino acid sequence identity to human CYP4B1, was cloned by polymerase chain reaction approaches. The results demonstrate the presence of wallaby CYP4B1 that shares several common features with other published CYP4Bs; however the wallaby CYP4B1 cDNA contains four extra amino acid residues at the NH2-terminal, a fundamentally conserved transmembrane anchor of all eukaryote CYPs.",
    keywords = "2 fluorenylamine, amino acid, complementary DNA, cytochrome P450, cytochrome P450 3A4, cytochrome P450 4B1, xenobiotic agent, aerobic metabolism, amino acid sequence, amino terminal sequence, animal tissue, article, Australia, CYP4B gene, CYP4B1 gene, eukaryote, gene, immunoreactivity, koala, liver microsome, marsupial, molecular cloning, nonhuman, nucleotide sequence, polymerase chain reaction, priority journal, protein expression, protein family, rabbit, sequence homology, signal transduction, species difference, tammar wallaby, Amino Acid Sequence, Animals, Aryl Hydrocarbon Hydroxylases, DNA, Complementary, Fluorenes, Immunoblotting, Kidney, Liver, Lung, Macropodidae, Microsomes, Molecular Sequence Data, Rabbits, Eukaryota, Macropus eugenii, Metatheria, Oryctolagus cuniculus, Phascolarctidae",
    author = "Milic, {Natalie L.} and Ngo, {Suong N.T.} and Marchant, {Ceilidh L.} and Height, {Tamara A.} and McKinnon, {Ross A.}",
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    Pulmonary cytochrome P450 enzymes belonging to the CYP4B subfamily from an Australian marsupial, the tammar wallaby (Macropus eugenii). / Milic, Natalie L.; Ngo, Suong N.T.; Marchant, Ceilidh L.; Height, Tamara A.; McKinnon, Ross A.

    In: Comparative Biochemistry and Physiology, Part C, Vol. 153, No. 1, 2011, p. 60-66.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Pulmonary cytochrome P450 enzymes belonging to the CYP4B subfamily from an Australian marsupial, the tammar wallaby (Macropus eugenii)

    AU - Milic, Natalie L.

    AU - Ngo, Suong N.T.

    AU - Marchant, Ceilidh L.

    AU - Height, Tamara A.

    AU - McKinnon, Ross A.

    PY - 2011

    Y1 - 2011

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    KW - liver microsome

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    KW - polymerase chain reaction

    KW - priority journal

    KW - protein expression

    KW - protein family

    KW - rabbit

    KW - sequence homology

    KW - signal transduction

    KW - species difference

    KW - tammar wallaby

    KW - Amino Acid Sequence

    KW - Animals

    KW - Aryl Hydrocarbon Hydroxylases

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    KW - Fluorenes

    KW - Immunoblotting

    KW - Kidney

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    KW - Microsomes

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    DO - 10.1016/j.cbpc.2010.08.006

    M3 - Article

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    EP - 66

    JO - Comparative Biochemistry and Physiology, Part C

    JF - Comparative Biochemistry and Physiology, Part C

    SN - 1532-0456

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    ER -