title: Arginine ADP-ribosylation mechanism based on structural snapshots of iota-toxin and actin complex.
abstract: Clostridium perfringens iota-toxin (Ia) mono-ADP ribosylates Arg177 of actin, leading to cytoskeletal disorganization and cell death. To fully understand the reaction mechanism of arginine-specific mono-ADP ribosyl transferase, the structure of the toxin-substrate protein complex must be characterized. Recently, we solved the crystal structure of Ia in complex with actin and the nonhydrolyzable NAD(+) analog βTAD (thiazole-4-carboxamide adenine dinucleotide); however, the structures of the NAD(+)-bound form (NAD(+)-Ia-actin) and the ADP ribosylated form [Ia-ADP ribosylated (ADPR)-actin] remain unclear. Accidentally, we found that ethylene glycol as cryo-protectant inhibits ADP ribosylation and crystallized the NAD(+)-Ia-actin complex. Here we report high-resolution structures of NAD(+)-Ia-actin and Ia-ADPR-actin obtained by soaking apo-Ia-actin crystal with NAD(+) under different conditions. The structures of NAD(+)-Ia-actin and Ia-ADPR-actin represent the pre- and postreaction states, respectively. By assigning the βTAD-Ia-actin structure to the transition state, the strain-alleviation model of ADP ribosylation, which we proposed previously, is experimentally confirmed and improved. Moreover, this reaction mechanism appears to be applicable not only to Ia but also to other ADP ribosyltransferases.
authors: Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-Motoyama, Kyoto 603-8555, Japan.
authors: Toshiharu
authors: T
authors: Tsurumura
authors: Yayoi
authors: Y
authors: Tsumori
authors: Hao
authors: H
authors: Qiu
authors: Masataka
authors: M
authors: Oda
authors: Jun
authors: J
authors: Sakurai
authors: Masahiro
authors: M
authors: Nagahama
authors: Hideaki
authors: H
authors: Tsuge
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry* / Q000378:metabolism; D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D001427:Bacterial Toxins / Q000737:chemistry* / Q000378:metabolism; D003016:Clostridium perfringens / Q000201:enzymology*; D009243:NAD / Q000031:analogs & derivatives / Q000737:chemistry* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary
chemical_list: D000199:Actins; D001427:Bacterial Toxins; C050737:iota toxin, Clostridium perfringens; D009243:NAD; D036002:ADP Ribose Transferases
references: Sci STKE. 2002 Dec 17;2002(163):pe53
references: 12488509
references: J Biol Chem. 1980 Nov 25;255(22):10717-20
references: 7000782
references: Methods Enzymol. 2003;374:492-509
references: 14696386
references: FEBS Lett. 1987 Feb 9;212(1):109-13
references: 3100333
references: J Biol Chem. 2003 Nov 14;278(46):45924-30
references: 12933793
references: Nat Struct Biol. 1999 Oct;6(10):932-6
references: 10504727
references: Nature. 1990 Sep 6;347(6288):44-9
references: 2395461
references: Nature. 1986 Jul 24-30;322(6077):390-2
references: 3736664
references: Proc Natl Acad Sci U S A. 1999 Aug 17;96(17):9459-64
references: 10449714
references: Annu Rev Biochem. 1985;54:73-100
references: 3927821
references: Science. 2010 Feb 26;327(5969):1139-42
references: 20185726
references: Proc Natl Acad Sci U S A. 1982 May;79(10):3129-33
references: 6954463
references: J Biol Chem. 2012 Oct 26;287(44):37030-41
references: 22969084
references: Protein Sci. 2008 May;17(5):878-86
references: 18369192
references: Proc Natl Acad Sci U S A. 2008 May 27;105(21):7399-404
references: 18490658
references: J Biol Chem. 2009 Oct 16;284(42):28713-9
references: 19692332
references: J Biol Chem. 1977 Apr 10;252(7):2455-7
references: 139409
references: J Mol Biol. 2006 Dec 8;364(4):705-15
references: 17027031
references: Proc Natl Acad Sci U S A. 1978 Jul;75(7):3050-4
references: 210449
references: Science. 2005 Aug 12;309(5737):1093-6
references: 16099990
references: J Mol Biol. 2001 Jan 5;305(1):95-107
references: 11114250
references: Amino Acids. 2011 Jul;41(2):257-69
references: 20652610
references: J Mol Biol. 2003 Jan 17;325(3):471-83
references: 12498797
references: Mol Microbiol. 1992 Oct;6(20):2905-8
references: 1479882
references: FEBS Lett. 2008 Apr 2;582(7):1032-6
references: 18325337
references: J Biol Chem. 2002 Jun 21;277(25):23028-36
references: 11948190
references: FEBS J. 2011 Dec;278(23):4526-43
references: 21466657
references: J Biol Chem. 2002 Aug 23;277(34):30950-7
references: 12029083
references: EMBO Rep. 2008 Aug;9(8):802-9
references: 18583986
references: FEBS Lett. 1996 Feb 19;380(3):291-5
references: 8601443
references: Structure. 2006 Aug;14(8):1219-29
references: 16905096
references: J Phys Chem B. 2011 Jun 23;115(24):7932-9
references: 21612228
references: Trends Biochem Sci. 2010 Apr;35(4):208-19
references: 20106667
references: J Mol Biol. 2002 Sep 27;322(4):687-96
references: 12270706
references: FEBS Lett. 1987 Dec 10;225(1-2):48-52
references: 2891567
references: J Biol Chem. 1979 Jul 25;254(14):6270-2
references: 221495
references: Nature. 2005 Aug 18;436(7053):979-84
references: 16107839
references: J Exp Med. 1969 Jan 1;129(1):1-21
references: 4304436