5WU0

Crystal structure of C. perfringens iota-like enterotoxin CPILE-a with NADH

5WU0 の概要

• エントリーDOI: 10.2210/pdb5wu0/pdb
• 関連するPDBエントリー: 5WTZ
• 分子名称: Binary enterotoxin of Clostridium perfringens component a, 1,4-DIHYDRONICOTINAMIDE ADENINE DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: bacterial toxin, actin, cpile-a, adp-ribosylation, toxin
• 由来する生物種: Clostridium perfringens
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 48336.87

構造登録者

Toniti, W.,Yoshida, T.,Tsurumura, T.,Irikura, D.,Tsuge, H. (登録日: 2016-12-15, 公開日: 2017-03-01, 最終更新日: 2023-11-08)

主引用文献

Toniti, W.,Yoshida, T.,Tsurumura, T.,Irikura, D.,Monma, C.,Kamata, Y.,Tsuge, H.
Crystal structure and structure-based mutagenesis of actin-specific ADP-ribosylating toxin CPILE-a as novel enterotoxin
PLoS ONE, 12:e0171278-e0171278, 2017

PubMed Abstract: Unusual outbreaks of food poisoning in Japan were reported in which Clostridium perfringens was strongly suspected to be the cause based on epidemiological information and fingerprinting of isolates. The isolated strains lack the typical C. perfringens enterotoxin (CPE) but secrete a new enterotoxin consisting of two components: C. perfringens iota-like enterotoxin-a (CPILE-a), which acts as an enzymatic ADP-ribosyltransferase, and CPILE-b, a membrane binding component. Here we present the crystal structures of apo-CPILE-a, NAD+-CPILE-a and NADH-CPILE-a. Though CPILE-a structure has high similarity with known iota toxin-a (Ia) with NAD+, it possesses two extra-long protruding loops from G262-S269 and E402-K408 that are distinct from Ia. Based on the Ia-actin complex structure, we focused on actin-binding interface regions (I-V) including two protruding loops (PT) and examined how mutations in these regions affect the ADP-ribosylation activity of CPILE-a. Though some site-directed mutagenesis studies have already been conducted on the actin binding site of Ia, in the present study, mutagenesis studies were conducted against both α- and β/γ-actin in CPILE-a and Ia. Interestingly, CPILE-a ADP-ribosylates both α- and β/γ-actin, but its sensitivity towards β/γ-actin is 36% compared with α-actin. Our results contrast to that only C2-I ADP-ribosylates β/γ-actin. We also showed that PT-I and two convex-concave interactions in CPILE-a are important for actin binding. The current study is the first detailed analysis of site-directed mutagenesis in the actin binding region of Ia and CPILE-a against both α- and β/γ-actin.

PubMed: 28199340
DOI: 10.1371/journal.pone.0171278

実験手法

X-RAY DIFFRACTION (2.251 Å)