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Visualization of actin mono‐ADP‐ribosylation in iota toxin and actin complex (555.4)
Author(s) -
Tsuge Hideaki,
Tsurumura Toshiharu,
Tsumori Yayoi
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.555.4
Subject(s) - nad+ kinase , adp ribosylation , actin , nicotinamide adenine dinucleotide , biology , chemistry , biophysics , biochemistry , enzyme
Clostridium perfringens iota‐toxin (Ia) mono‐ADP‐ribosylates Arg177 of actin, leading to cytoskeletal disorganization and cell death. We revealed the crystal structure of Ia in complex with actin and the nonhydrolyzable NAD+ analog βTAD (thiazole‐4‐carboxamide adenine dinucleotide) (ref1); however, the structures of the NAD+ bound form (NAD+‐Ia‐actin) and the ADP ribosylated form [Ia‐ADP ribosylated (ADPR)‐actin] remain unclear. Here we obtained the good diffraction apo crystals and found that ethylene glycol as cryo‐protectant inhibits ADP ribosylation. We used the apo crystal as the ADP‐ribosylation reaction chamber and applied NAD+‐soaking to this complex in different conditions. Finally, we obtained high‐resolution structures of NAD+‐Ia‐actin and Ia‐ADPR‐actin (ref2). The structures of NAD+‐Ia‐actin and Ia‐ADPR‐actin represent the pre‐ and postreaction states, respectively. The strain‐alleviation model of ADP ribosylation, which we proposed previously, became more plausible with additional snapshots of actin ADP‐ribosylation. (1) Tsuge H, Nagahama M, Oda M, Iwamoto S, Utsunomiya H, Marquez VE, Katunuma N, Nishizawa M, Sakurai J (2008) PNAS 105 (21):7399‐7404. (2) Tsurumura T, Tsumori Y, Qiu H, Oda M, Sakurai J, Nagahama M, Tsuge H (2013) PNAS 110 (11):4267‐4272.