DOI: 10.1073/pnas.0801215105. Structural basis of actin recognition and arginine ADP-ribosylation by <i>Clostridium perfringens</i> ι-toxin

Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens ι-toxin

10.1073/pnas.0801215105

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The ADP-ribosylating toxins (ADPRTs) produced by pathogenic bacteria modify intracellular protein and affect eukaryotic cell function. Actin-specific ADPRTs (including Clostridium perfringens ι-toxin and Clostridium botulinum C2 toxin) ADP-ribosylate G-actin at Arg-177, leading to disorganization of the cytoskeleton and cell death. Although the structures of many actin-specific ADPRTs are available, the mechanisms underlying actin recognition and selective ADP-ribosylation of Arg-177 remain unknown. Here we report the crystal structure of actin-Ia in complex with the nonhydrolyzable NAD analog βTAD at 2.8 Å resolution. The structure indicates that Ia recognizes actin via five loops around NAD: loop I (Tyr-60–Tyr-62 in the N domain), loop II (active-site loop), loop III, loop IV (PN loop), and loop V (ADP-ribosylating turn–turn loop). We used site-directed mutagenesis to confirm that loop I on the N domain and loop II are essential for the ADP-ribosyltransferase activity. Furthermore, we revealed that Glu-378 on the EXE loop is in close proximity to Arg-177 in actin, and we proposed that the ADP-ribosylation of Arg-177 proceeds by an SN1 reaction via first an oxocarbenium ion intermediate and second a cationic intermediate by alleviating the strained conformation of the first oxocarbenium ion. Our results suggest a common reaction mechanism for ADPRTs. Moreover, the structure might be of use in rational drug design to block toxin-substrate recognition.

Bibliography

Tsuge, H., Nagahama, M., Oda, M., Iwamoto, S., Utsunomiya, H., Marquez, V. E., Katunuma, N., Nishizawa, M., & Sakurai, J. (2008). Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens Î¹-toxin. Proceedings of the National Academy of Sciences, 105(21), 7399â7404.

Authors 9

Hideaki Tsuge (first)
Masahiro Nagahama (additional)
Masataka Oda (additional)
Shinobu Iwamoto (additional)
Hiroko Utsunomiya (additional)
Victor E. Marquez (additional)
Nobuhiko Katunuma (additional)
Mugio Nishizawa (additional)
Jun Sakurai (additional)

References 38 Referenced 101

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WL DeLano The PyMOL Users Manual (DeLano Scientific, Palo Alto, CA, 2002). / The PyMOL Users Manual by DeLano WL (2002)

Dates

Type	When
Created	17 years, 3 months ago (May 19, 2008, 8:58 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 4:47 p.m.)
Indexed	4 weeks, 2 days ago (July 30, 2025, 11:20 a.m.)
Issued	17 years, 3 months ago (May 27, 2008)
Published	17 years, 3 months ago (May 27, 2008)
Published Online	17 years, 3 months ago (May 27, 2008)
Published Print	17 years, 3 months ago (May 27, 2008)

Funders 0

None

BibTeX

@article{Tsuge_2008, title={Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens ι-toxin}, volume={105}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0801215105}, DOI={10.1073/pnas.0801215105}, number={21}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Tsuge, Hideaki and Nagahama, Masahiro and Oda, Masataka and Iwamoto, Shinobu and Utsunomiya, Hiroko and Marquez, Victor E. and Katunuma, Nobuhiko and Nishizawa, Mugio and Sakurai, Jun}, year={2008}, month=may, pages={7399–7404} }

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  "abstract": "<jats:p>\n            The ADP-ribosylating toxins (ADPRTs) produced by pathogenic bacteria modify intracellular protein and affect eukaryotic cell function. Actin-specific ADPRTs (including\n            <jats:italic>Clostridium perfringens</jats:italic>\n            \u03b9-toxin and\n            <jats:italic>Clostridium botulinum</jats:italic>\n            C2 toxin) ADP-ribosylate G-actin at Arg-177, leading to disorganization of the cytoskeleton and cell death. Although the structures of many actin-specific ADPRTs are available, the mechanisms underlying actin recognition and selective ADP-ribosylation of Arg-177 remain unknown. Here we report the crystal structure of actin-Ia in complex with the nonhydrolyzable NAD analog \u03b2TAD at 2.8 \u212b resolution. The structure indicates that Ia recognizes actin via five loops around NAD: loop I (Tyr-60\u2013Tyr-62 in the N domain), loop II (active-site loop), loop III, loop IV (PN loop), and loop V (ADP-ribosylating turn\u2013turn loop). We used site-directed mutagenesis to confirm that loop I on the N domain and loop II are essential for the ADP-ribosyltransferase activity. Furthermore, we revealed that Glu-378 on the EXE loop is in close proximity to Arg-177 in actin, and we proposed that the ADP-ribosylation of Arg-177 proceeds by an SN1 reaction via first an oxocarbenium ion intermediate and second a cationic intermediate by alleviating the strained conformation of the first oxocarbenium ion. Our results suggest a common reaction mechanism for ADPRTs. Moreover, the structure might be of use in rational drug design to block toxin-substrate recognition.\n          </jats:p>",
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