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Related Experiment Videos

Cytochrome c specific methylase from wheat germ.

P DiMaria, S Kim, W K Paik

    Biochemistry
    |March 2, 1982
    PubMed
    Summary

    Wheat germ cytochrome c methylase specifically methylates lysine 72 in vitro, not residue 86. This enzyme prefers avian cytochromes c and may methylate apocytochrome c before heme attachment.

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    Area of Science:

    • Biochemistry
    • Plant molecular biology
    • Protein modification

    Background:

    • Plant cytochromes c contain trimethyllysines at residues 72 and 86.
    • Understanding the specific methylation sites and enzymes involved is crucial for elucidating protein function.

    Purpose of the Study:

    • To identify the specific site of in vitro methylation of cytochrome c by a wheat germ enzyme.
    • To characterize the substrate specificity and kinetic properties of the purified methyltransferase.
    • To investigate potential inhibitors of the enzyme.

    Main Methods:

    • Purification of cytochrome c-specific N-methyltransferase (protein methylase III) from wheat germ.
    • Two-dimensional peptide mapping, Aminex A-5 column peptide analysis, and CNBr cleavage analysis.
    • In vitro methylation assays using various cytochrome c forms and S-adenosyl-L-homocysteine analogues.

    Main Results:

    • The purified enzyme and crude extract methylated lysine 72 of horse heart cytochrome c in vitro.
    • Residue 86 was not methylated, and avian cytochromes c were better substrates than mammalian ones.
    • The enzyme exhibited a low Km for apocytochrome c, and S-adenosylhomocysteine acted as an inhibitor.

    Conclusions:

    • Wheat germ cytochrome c methylase specifically targets lysine 72.
    • Methylation likely occurs prior to heme attachment, with avian cytochromes c being preferred substrates.
    • Methylated lysine residues decrease peptide basicity, affecting chromatographic behavior.

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