Crystal structures of OrfX1, OrfX2 and the OrfX1–OrfX3 complex from the orfX gene cluster of botulinum neurotoxin E1

Gao, Linfeng; Lam, Kwok‐ho; Liu, Shun; Przykopanski, Adina; Lübke, Johanna; Qi, Ruifeng; Krüger, Maren; Nowakowska, Maria B.; Selby, Katja; Douillard, François P.; Dorner, Martin B.; Perry, Kay; Lindström, Miia; Dorner, Brigitte G.; Rummel, Andreas; Jin, Rongsheng

doi:10.1002/1873-3468.14576

Crystal structures of OrfX1, OrfX2 and the OrfX1–OrfX3 complex from the orfX gene cluster of botulinum neurotoxin E1

Journal Article · Wed Jan 18 00:00:00 EST 2023 · FEBS Letters

DOI:https://doi.org/10.1002/1873-3468.14576· OSTI ID:2423355

Gao, Linfeng ^[1]; Lam, Kwok‐ho ^[1]; Liu, Shun ^[1]; Przykopanski, Adina ^[2]; Lübke, Johanna ^[2]; Qi, Ruifeng ^[1]; Krüger, Maren ^[3]; ^[4]; Selby, Katja ^[4]; Douillard, François P. ^[4]; Dorner, Martin B. ^[3]; Perry, Kay ^[5]; Lindström, Miia ^[4]; Dorner, Brigitte G. ^[3]; Rummel, Andreas ^[2]; ^[1]

University of California, Irvine, CA (United States)
Hannover Medical School (Germany)
Robert Koch Institute, Berlin (Germany)
University of Helsinki (Finland)
Argonne National Laboratory (ANL), Argonne, IL (United States)

Botulinum neurotoxins (BoNTs) are among the most lethal toxins known to humans, comprising seven established serotypes termed BoNT/A–G encoded in two types of gene clusters (ha and orfX) in BoNT-producing clostridia. The ha cluster encodes four non-toxic neurotoxin-associated proteins (NAPs) that assemble with BoNTs to protect and enhance their oral toxicity. However, the structure and function of the orfX-type NAPs remain largely unknown. Here, we report the crystal structures for OrfX1, OrfX2, and an OrfX1–OrfX3 complex, which are encoded in the orfX cluster of a BoNT/E1-producing Clostridium botulinum strain associated with human foodborne botulism. These structures lay the foundation for future studies on the potential roles of OrfX proteins in oral intoxication and pathogenesis of BoNTs.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: Deutsche Forschungsgemeinschaft; European Research Council; National Institute of Allergy and Infectious Diseases (NIAID); National Institutes of Health (NIH); Swiss Federal Office for Civil Protection; USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 2423355

Journal Information:: FEBS Letters, Journal Name: FEBS Letters Journal Issue: 4 Vol. 597; ISSN 0014-5793

Publisher:: Federation of European Biochemical SocietiesCopyright Statement

Country of Publication:: United States

Language:: English

References (60)

Crystal structures of OrfX2 and P47 from a Botulinum neurotoxin OrfX-type gene cluster Gustafsson, Robert; Berntsson, Ronnie P-A; Martínez-Carranza, Markel FEBS Letters, Vol. 591, Issue 22 https://doi.org/10.1002/1873-3468.12889	journal	November 2017
Identification of a novel botulinum neurotoxin gene cluster in Enterococcus Brunt, Jason; Carter, Andrew T.; Stringer, Sandra C. FEBS Letters, Vol. 592, Issue 3 https://doi.org/10.1002/1873-3468.12969	journal	January 2018
Atomic Structures of the Human Immunophilin FKBP-12 Complexes with FK506 and Rapamycin Van Duyne, Gregory D.; Standaert, Robert F.; Karplus, P. Andrew Journal of Molecular Biology, Vol. 229, Issue 1 https://doi.org/10.1006/jmbi.1993.1012	journal	January 1993
Conserved structure of genes encoding components of botulinum neurotoxin complex M and the sequence of the gene coding for the nontoxic component in nonproteolyticClostridium botulinum type F East, Alison K.; Collins, Matthew D. Current Microbiology, Vol. 29, Issue 2 https://doi.org/10.1007/BF01575751	journal	August 1994
Analysis of the Botulinum Neurotoxin Type F Gene Clusters in Proteolytic and Nonproteolytic Clostridium botulinum and Clostridium barati East, Alison K.; Bhandari, Manju; Hielm, Sebastian Current Microbiology, Vol. 37, Issue 4 https://doi.org/10.1007/s002849900376	journal	October 1998
The three-dimensional structure of human bactericidal/permeability-increasing protein Beamer, Lesa J.; Carroll, Stephen F.; Eisenberg, David Biochemical Pharmacology, Vol. 57, Issue 3 https://doi.org/10.1016/S0006-2952(98)00279-2	journal	February 1999
Regulation of neurotoxin complex expression in Clostridium botulinum strains 62A, Hall A-hyper, and NCTC 2916 Bradshaw, Marite; Dineen, Sean S.; Maks, Nicole D. Anaerobe, Vol. 10, Issue 6 https://doi.org/10.1016/j.anaerobe.2004.07.001	journal	December 2004
Binding properties of Clostridium botulinum type C progenitor toxin to mucins Nakamura, Toshio; Takada, Noriko; Tonozuka, Takashi Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1770, Issue 4 https://doi.org/10.1016/j.bbagen.2006.11.006	journal	April 2007
The TULIP superfamily of eukaryotic lipid-binding proteins as a mediator of lipid sensing and transport Alva, Vikram; Lupas, Andrei N. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1861, Issue 8 https://doi.org/10.1016/j.bbalip.2016.01.016	journal	August 2016
Clostridium botulinum type A progenitor toxin binds to Intestine-407 cells via N-acetyllactosamine moiety Kojima, Shoudou; Eguchi, Hironobu; Ookawara, Tomomi Biochemical and Biophysical Research Communications, Vol. 331, Issue 2 https://doi.org/10.1016/j.bbrc.2005.04.007	journal	June 2005
Small-angle X-ray scattering reveals structural dynamics of the botulinum neurotoxin associating protein, nontoxic nonhemagglutinin Sagane, Yoshimasa; Miyashita, Shin-Ichiro; Miyata, Keita Biochemical and Biophysical Research Communications, Vol. 425, Issue 2 https://doi.org/10.1016/j.bbrc.2012.07.077	journal	August 2012
High-resolution crystal structure of HA33 of botulinum neurotoxin type B progenitor toxin complex Lee, Kwangkook; Lam, Kwok-Ho; Kruel, Anna Magdalena Biochemical and Biophysical Research Communications, Vol. 446, Issue 2 https://doi.org/10.1016/j.bbrc.2014.03.008	journal	April 2014
Identification of a Botulinum Neurotoxin-like Toxin in a Commensal Strain of Enterococcus faecium Zhang, Sicai; Lebreton, Francois; Mansfield, Michael J. Cell Host & Microbe, Vol. 23, Issue 2 https://doi.org/10.1016/j.chom.2017.12.018	journal	February 2018
Safety and pharmacodynamics of a novel recombinant botulinum toxin E (rBoNT-E): Results of a phase 1 study in healthy male subjects compared with abobotulinumtoxinA (Dysport®) Pons, Laurent; Vilain, Claire; Volteau, Magali Journal of the Neurological Sciences, Vol. 407 https://doi.org/10.1016/j.jns.2019.116516	journal	December 2019
Architecture of the botulinum neurotoxin complex: a molecular machine for protection and delivery Lam, Kwok-Ho; Jin, Rongsheng Current Opinion in Structural Biology, Vol. 31 https://doi.org/10.1016/j.sbi.2015.03.013	journal	April 2015
Rational Protein Crystallization by Mutational Surface Engineering Derewenda, Zygmunt S. Structure, Vol. 12, Issue 4 https://doi.org/10.1016/j.str.2004.03.008	journal	April 2004
The hypothetical protein P47 of Clostridium botulinum E1 strain Beluga has a structural topology similar to bactericidal/permeability-increasing protein Lam, Kwok-ho; Qi, Ruifeng; Liu, Shun Toxicon, Vol. 147 https://doi.org/10.1016/j.toxicon.2017.10.012	journal	June 2018
Structure and function of tetanus and botulinum neurotoxins Montecucco, Cesare; Schiavo, Giampietro Quarterly Reviews of Biophysics, Vol. 28, Issue 4 https://doi.org/10.1017/S0033583500003292	journal	November 1995
Protein Fusions of BPI with CETP Retain Functions Inherent to Each Lloyd, David B.; Bonnette, Peter; Thompson, John F. Biochemistry, Vol. 45, Issue 43 https://doi.org/10.1021/bi0615590	journal	October 2006
Free R value: a novel statistical quantity for assessing the accuracy of crystal structures Brünger, Axel T. Nature, Vol. 355, Issue 6359 https://doi.org/10.1038/355472a0	journal	January 1992
Identification and characterization of a novel botulinum neurotoxin Zhang, Sicai; Masuyer, Geoffrey; Zhang, Jie Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/ncomms14130	journal	August 2017
Botulinum neurotoxins: genetic, structural and mechanistic insights Rossetto, Ornella; Pirazzini, Marco; Montecucco, Cesare Nature Reviews Microbiology, Vol. 12, Issue 8 https://doi.org/10.1038/nrmicro3295	journal	June 2014
A neurotoxin that specifically targets Anopheles mosquitoes Contreras, Estefania; Masuyer, Geoffrey; Qureshi, Nadia Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-019-10732-w	journal	June 2019
Molecular Assembly of Clostridium botulinum progenitor M complex of type E Eswaramoorthy, Subramaniam; Sun, Jingchuan; Li, Huilin Scientific Reports, Vol. 5, Issue 1 https://doi.org/10.1038/srep17795	journal	December 2015
Structure of human BPI (bactericidal/permeability-increasing protein) and implications for related proteins Beamer, L. J. Biochemical Society Transactions, Vol. 31, Issue 4 https://doi.org/10.1042/BST0310791	journal	August 2003
Bioinformatics of the TULIP domain superfamily Kopec, Klaus O.; Alva, Vikram; Lupas, Andrei N. Biochemical Society Transactions, Vol. 39, Issue 4 https://doi.org/10.1042/BST0391033	journal	July 2011
Molecular assembly of botulinum neurotoxin progenitor complexes Benefield, D. A.; Dessain, S. K.; Shine, N. Proceedings of the National Academy of Sciences, Vol. 110, Issue 14 https://doi.org/10.1073/pnas.1222139110	journal	March 2013
Conserved SMP domains of the ERMES complex bind phospholipids and mediate tether assembly AhYoung, Andrew P.; Jiang, Jiansen; Zhang, Jiang Proceedings of the National Academy of Sciences, Vol. 112, Issue 25 https://doi.org/10.1073/pnas.1422363112	journal	June 2015
Crystal Structure of Clostridium botulinum Whole Hemagglutinin Reveals a Huge Triskelion-shaped Molecular Complex Amatsu, Sho; Sugawara, Yo; Matsumura, Takuhiro Journal of Biological Chemistry, Vol. 288, Issue 49 https://doi.org/10.1074/jbc.M113.521179	journal	December 2013
Botulinum hemagglutinin disrupts the intercellular epithelial barrier by directly binding E-cadherin Sugawara, Yo; Matsumura, Takuhiro; Takegahara, Yuki Journal of Cell Biology, Vol. 189, Issue 4 https://doi.org/10.1083/jcb.200910119	journal	May 2010
Structural and biochemical characterization of the protease domain of the mosaic botulinum neurotoxin type HA Lam, Kwok-ho; Sikorra, Stefan; Weisemann, Jasmin Pathogens and Disease, Vol. 76, Issue 4 https://doi.org/10.1093/femspd/fty044	journal	April 2018
Molecular Characterization of a Novel Botulinum Neurotoxin Type H Gene Dover, Nir; Barash, Jason R.; Hill, Karen K. The Journal of Infectious Diseases, Vol. 209, Issue 2 https://doi.org/10.1093/infdis/jit450	journal	October 2013
Search and sequence analysis tools services from EMBL-EBI in 2022 Madeira, Fábio; Pearce, Matt; Tivey, Adrian R. N. Nucleic Acids Research, Vol. 50, Issue W1 https://doi.org/10.1093/nar/gkac240	journal	April 2022
Deciphering key features in protein structures with the new ENDscript server Robert, Xavier; Gouet, Patrice Nucleic Acids Research, Vol. 42, Issue W1 https://doi.org/10.1093/nar/gku316	journal	April 2014
Safety and Efficacy of EB-001, a Novel Type E Botulinum Toxin, in Subjects with Glabellar Frown Lines: Results of a Phase 2, Randomized, Placebo-Controlled, Ascending-Dose Study Yoelin, Steve G.; Dhawan, Sunil S.; Vitarella, Domenico Plastic & Reconstructive Surgery, Vol. 142, Issue 6 https://doi.org/10.1097/PRS.0000000000005029	journal	December 2018
The complete nucleotide sequence of the gene encoding the nontoxic component of Clostridium botulinum type E progenitor toxin Fujii, N.; Kimura, K.; Yokosawa, N. Journal of General Microbiology, Vol. 139, Issue 1 https://doi.org/10.1099/00221287-139-1-79	journal	January 1993
The haemagglutinin of Clostridium botulinum type C progenitor toxin plays an essential role in binding of toxin to the epithelial cells of guinea pig small intestine, leading to the efficient absorption of the toxin Fujinaga, Yukako; Inoue, Kaoru; Watanabe, Sadahiro Microbiology, Vol. 143, Issue 12 https://doi.org/10.1099/00221287-143-12-3841	journal	December 1997
Clostridium botulinum type A haemagglutinin-positive progenitor toxin (HA+-PTX) binds to oligosaccharides containing Galβ1-4GlcNAc through one subcomponent of haemagglutinin (HA1) Inoue, Kaoru; Fujinaga, Yukako; Honke, Koichi Microbiology, Vol. 147, Issue 4 https://doi.org/10.1099/00221287-147-4-811	journal	April 2001
Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal epithelial cells and erythrocytes Fujinaga, Yukako; Inoue, Kaoru; Watarai, Shinobu Microbiology, Vol. 150, Issue 5 https://doi.org/10.1099/mic.0.26805-0	journal	May 2004
Phaser crystallographic software McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D. Journal of Applied Crystallography, Vol. 40, Issue 4 https://doi.org/10.1107/S0021889807021206	journal	July 2007
Coot model-building tools for molecular graphics Emsley, Paul; Cowtan, Kevin Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132 https://doi.org/10.1107/S0907444904019158	journal	November 2004
MolProbity : all-atom structure validation for macromolecular crystallography Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1 https://doi.org/10.1107/S0907444909042073	journal	December 2009
XDS Kabsch, Wolfgang Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2 https://doi.org/10.1107/S0907444909047337	journal	January 2010
PHENIX: a comprehensive Python-based system for macromolecular structure solution Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221 https://doi.org/10.1107/S0907444909052925	journal	January 2010
Toward rational protein crystallization: A Web server for the design of crystallizable protein variants Goldschmidt, Lukasz; Cooper, David R.; Derewenda, Zygmunt S. Protein Science, Vol. 16, Issue 8 https://doi.org/10.1110/ps.072914007	journal	August 2007
The HA proteins of botulinum toxin disrupt intestinal epithelial intercellular junctions to increase toxin absorption Matsumura, Takuhiro; Jin, Yingji; Kabumoto, Yuko Cellular Microbiology, Vol. 0, Issue 0 https://doi.org/10.1111/j.1462-5822.2007.01048.x	journal	September 2007
Botulinum Neurotoxin Is Shielded by NTNHA in an Interlocked Complex Gu, S.; Rumpel, S.; Zhou, J. Science, Vol. 335, Issue 6071 https://doi.org/10.1126/science.1214270	journal	February 2012
Molecular basis for disruption of E-cadherin adhesion by botulinum neurotoxin A complex Lee, K.; Zhong, X.; Gu, S. Science, Vol. 344, Issue 6190 https://doi.org/10.1126/science.1253823	journal	June 2014
Quantitative Real-Time Reverse Transcription-PCR Analysis Reveals Stable and Prolonged Neurotoxin Cluster Gene Activity in a Clostridium botulinum Type E Strain at Refrigeration Temperature Chen, Ying; Korkeala, Hannu; Lindén, Jere Applied and Environmental Microbiology, Vol. 74, Issue 19 https://doi.org/10.1128/AEM.00469-08	journal	October 2008
Laboratory Diagnostics of Botulism Lindström, Miia; Korkeala, Hannu Clinical Microbiology Reviews, Vol. 19, Issue 2 https://doi.org/10.1128/CMR.19.2.298-314.2006	journal	April 2006
Toxigenic clostridia. Hatheway, C. L. Clinical Microbiology Reviews, Vol. 3, Issue 1 https://doi.org/10.1128/CMR.3.1.66	journal	January 1990
Clostridium botulinum neurotoxin. Sugiyama, H. Microbiological Reviews, Vol. 44, Issue 3 https://doi.org/10.1128/MMBR.44.3.419-448.1980	journal	January 1980
Recombination and insertion events involving the botulinum neurotoxin complex genes in Clostridium botulinum types A, B, E and F and Clostridium butyricumtype E strains Hill, Karen K.; Xie, Gary; Foley, Brian T. BMC Biology, Vol. 7, Issue 1 https://doi.org/10.1186/1741-7007-7-66	journal	October 2009
Functional Dissection of the Clostridium botulinum Type B Hemagglutinin Complex: Identification of the Carbohydrate and E-Cadherin Binding Sites Sugawara, Yo; Yutani, Masahiro; Amatsu, Sho PLoS ONE, Vol. 9, Issue 10 https://doi.org/10.1371/journal.pone.0111170	journal	October 2014
Structure of a Bimodular Botulinum Neurotoxin Complex Provides Insights into Its Oral Toxicity Lee, Kwangkook; Gu, Shenyan; Jin, Lei PLoS Pathogens, Vol. 9, Issue 10 https://doi.org/10.1371/journal.ppat.1003690	journal	October 2013
Looking for the X Factor in Bacterial Pathogenesis: Association of orfX-p47 Gene Clusters with Toxin Genes in Clostridial and Non-Clostridial Bacterial Species Nowakowska, Maria B.; Douillard, François P.; Lindström, Miia Toxins, Vol. 12, Issue 1 https://doi.org/10.3390/toxins12010019	journal	December 2019
Human Botulism in France, 1875–2016 Rasetti-Escargueil, Christine; Lemichez, Emmanuel; Popoff, Michel R. Toxins, Vol. 12, Issue 5 https://doi.org/10.3390/toxins12050338	journal	May 2020
Toxemia in Human Naturally Acquired Botulism Rasetti-Escargueil, Christine; Lemichez, Emmanuel; Popoff, Michel R. Toxins, Vol. 12, Issue 11 https://doi.org/10.3390/toxins12110716	journal	November 2020
Crystal Structure of the Receptor-Binding Domain of Botulinum Neurotoxin Type HA, Also Known as Type FA or H Yao, Guorui; Lam, Kwok-ho; Perry, Kay Toxins, Vol. 9, Issue 3 https://doi.org/10.3390/toxins9030093	journal	March 2017
Characterization of Hemagglutinin Negative Botulinum Progenitor Toxins Kalb, Suzanne; Baudys, Jakub; Smith, Theresa Toxins, Vol. 9, Issue 6 https://doi.org/10.3390/toxins9060193	journal	June 2017

Similar Records

The hypothetical protein P47 of Clostridium botulinum E1 strain Beluga has a structural topology similar to bactericidal/permeability-increasing protein

Journal Article · Sun Oct 15 20:00:00 EDT 2017 · Toxicon · OSTI ID:1434740

Differentiating Botulinum Neurotoxin-Producing Clostridia with a Simple, Multiplex PCR Assay

Journal Article · Thu Jul 20 20:00:00 EDT 2017 · Applied and Environmental Microbiology · OSTI ID:1626064

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
OrfX1
OrfX2
OrfX3
bacterial toxin
biochemistry
biophysics
botulinum neurotoxin
cell biology
molecular biology
orfX gene cluster

Crystal structures of OrfX1, OrfX2 and the OrfX1–OrfX3 complex from the orfX gene cluster of botulinum neurotoxin E1

Citation Formats

References (60)

Similar Records

Related Subjects