Activation of the protease from human adenovirus type 2 is accompanied by a conformational change that is dependent on cysteine-104

Sarah J. Jones, Munir Iqbal, Alasdair W. Grierson, Graham Kemp

    Research output: Contribution to journalArticle

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    Abstract

    Adenovirus codes for a protease the activity of which can be regulated in vitro by an 11 residue peptide (GVQSLKRRRCF) derived from another viral protein, pVI. Three cysteine residues, one in the activating peptide and two in the protease (C104 and C122), play a central role in both activation and catalysis. Expression of protease mutants in insect cells has shown that C104 is not essential for proteolytic activity. GVQSLKRRRCF also caused a concentration-dependent increase in tryptophan fluorescence of protease expressed in Escherichia coli that paralleled the increase in proteolytic activity, indicating that activation was accompanied by a conformational change. Tryptophan fluorescence of C104S was not increased by the addition of GVQSLKRRRCF, nor was the fluorescence of wild-type protease increased by the addition of the peptide analogues where cysteine is replaced by aspartic acid or serine, suggesting that C104 is involved in activation and C122 in catalysis.
    Original languageEnglish
    Pages (from-to)1821-1824
    JournalJournal of General Virology
    Volume77
    Issue number8
    DOIs
    Publication statusPublished - Aug 1996

    Fingerprint

    Human Adenoviruses
    Cysteine
    Peptide Hydrolases
    Fluorescence
    Catalysis
    Tryptophan
    Peptides
    Viral Proteins
    Adenoviridae
    Aspartic Acid
    Serine
    Insects
    Escherichia coli

    Cite this

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    title = "Activation of the protease from human adenovirus type 2 is accompanied by a conformational change that is dependent on cysteine-104",
    abstract = "Adenovirus codes for a protease the activity of which can be regulated in vitro by an 11 residue peptide (GVQSLKRRRCF) derived from another viral protein, pVI. Three cysteine residues, one in the activating peptide and two in the protease (C104 and C122), play a central role in both activation and catalysis. Expression of protease mutants in insect cells has shown that C104 is not essential for proteolytic activity. GVQSLKRRRCF also caused a concentration-dependent increase in tryptophan fluorescence of protease expressed in Escherichia coli that paralleled the increase in proteolytic activity, indicating that activation was accompanied by a conformational change. Tryptophan fluorescence of C104S was not increased by the addition of GVQSLKRRRCF, nor was the fluorescence of wild-type protease increased by the addition of the peptide analogues where cysteine is replaced by aspartic acid or serine, suggesting that C104 is involved in activation and C122 in catalysis.",
    author = "Jones, {Sarah J.} and Munir Iqbal and Grierson, {Alasdair W.} and Graham Kemp",
    year = "1996",
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    doi = "10.1099/0022-1317-77-8-1821",
    language = "English",
    volume = "77",
    pages = "1821--1824",
    journal = "Journal of General Virology",
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    Activation of the protease from human adenovirus type 2 is accompanied by a conformational change that is dependent on cysteine-104. / Jones, Sarah J.; Iqbal, Munir; Grierson, Alasdair W.; Kemp, Graham.

    In: Journal of General Virology, Vol. 77, No. 8, 08.1996, p. 1821-1824.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Activation of the protease from human adenovirus type 2 is accompanied by a conformational change that is dependent on cysteine-104

    AU - Jones, Sarah J.

    AU - Iqbal, Munir

    AU - Grierson, Alasdair W.

    AU - Kemp, Graham

    PY - 1996/8

    Y1 - 1996/8

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    AB - Adenovirus codes for a protease the activity of which can be regulated in vitro by an 11 residue peptide (GVQSLKRRRCF) derived from another viral protein, pVI. Three cysteine residues, one in the activating peptide and two in the protease (C104 and C122), play a central role in both activation and catalysis. Expression of protease mutants in insect cells has shown that C104 is not essential for proteolytic activity. GVQSLKRRRCF also caused a concentration-dependent increase in tryptophan fluorescence of protease expressed in Escherichia coli that paralleled the increase in proteolytic activity, indicating that activation was accompanied by a conformational change. Tryptophan fluorescence of C104S was not increased by the addition of GVQSLKRRRCF, nor was the fluorescence of wild-type protease increased by the addition of the peptide analogues where cysteine is replaced by aspartic acid or serine, suggesting that C104 is involved in activation and C122 in catalysis.

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