Identification of Glu173 as the critical amino acid residue for the ADP-ribosyltransferase activity of Clostridium botulinum C3 exoenzyme

Yuji Saito, Yasuo Nemoto, Toshimasa Ishizaki, Naoki Watanabe, Narito Morii, Shuh Narumiya

    Research output: Contribution to journalArticlepeer-review

    17 Citations (Scopus)

    Abstract

    Clostridium botulinum C3 exoenzyme specifically ADP-ribosylates rho-p21 in eukaryotic cells. Trp18 and Glu173 of this enzyme were substituted with other amino acids via site-directed mutagenesis. All substitutions at Glu173 caused a significant reduction in affinity for NAD and diminished ADP-ribosyltransferase activity. On the other hand, the activity of enzymes with the substitution at Trp18 remained intact. Swiss 3T3 cells treated with the enzyme with the Trp18 substitution showed the typical morphologic changes of the C3 exoenzyme phenotype. In contrast, no changes were found in cells incubated with the Glu173-substituted enzyme. These results indicate that the Glu173 residue of the C3 exoenzyme plays a key role in interacting with NAD and in expression of ADP-ribosyltransferase activity, which is essential for the phenotypic change by C3 exoenzyme treatment.

    Original languageEnglish
    Pages (from-to)105-109
    Number of pages5
    JournalFEBS Letters
    Volume371
    Issue number2
    DOIs
    Publication statusPublished - 1995 Sep 4

    Keywords

    • ADP-ribosylation
    • Botulinum C3 exoenzyme
    • Swiss 3T3 cell
    • rho p21

    ASJC Scopus subject areas

    • Biophysics
    • Structural Biology
    • Biochemistry
    • Molecular Biology
    • Genetics
    • Cell Biology

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