Partial purification of protein farnesyl cysteine carboxyl methyltransferase from bovine brain

Byung Cheol Yoo, Myung Seo Kang, Sangduk Kim, Young Sun Lee, Soo Yeon Choi, Chong Keun Ryu, Gil Hong Park, Jong Seol Han

    Research output: Contribution to journalArticlepeer-review

    10 Citations (Scopus)


    C-terminal farnesyl cysteine carboxyl methylation has been known to be the last step in the post-translational modification processes of several important signal transduction proteins in eukaryotes including ras related GTP binding proteins and the γ-subunit of heterotrimeric G proteins. Protein farnesyl cysteine carboxyl methyltransferase (PFCCMT; EC, catalyzing the reaction is well characterized as being stimulated by guanosine 5'-O-(3-thiotriphosphate) (GTPγS) and suppressed by N-acetyl-S-farnesyl-L-cysteine (AFC). As an initial step to understand the physiological significance of the process, we attempted to purify the enzyme, which was partially purified 130-fold (specific activity, 143 pmol of methyl group transferred/min/mg of protein) with yield of 1.8% after purification by fast protein liquid chromatography (FPLC) on a Superdex 75 column. The enzyme was further purified with non denaturing polyacrylamide gel electrophoresis (ND-PAGE) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of PFCCMT was determined to be about 30 kDa based on Superdex 75 FPLC as well as photoaffinity labelling with S-adenosyl-L-[methyl-3H] methionine ([methyl-3H]SAM). The partially purified enzyme (Superdex 75 eluate) was found to be characteristically affected by GTPγS, being activated about 40-fold in 2 mM, in contrast to ATP which did not show any effect on enzyme activity. Meanwhile, the enzyme was found to be markedly inhibited by AFC, reaching 0 activity in 2 mM. These observations strongly suggested that the partially purified enzyme was PFCCMT.

    Original languageEnglish
    Pages (from-to)227-234
    Number of pages8
    JournalExperimental and Molecular Medicine
    Issue number4
    Publication statusPublished - 1998 Dec 31


    • AFC
    • G-proteins
    • GTPγS
    • Partial purification of farnesylated cysteine carboxyl methyltransferase

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Medicine
    • Molecular Biology
    • Clinical Biochemistry


    Dive into the research topics of 'Partial purification of protein farnesyl cysteine carboxyl methyltransferase from bovine brain'. Together they form a unique fingerprint.

    Cite this