Structural Features of Neurotoxin Subtype A2 Cell Binding Domain

Overview

Journal Toxins (Basel)

Publisher MDPI

Specialty Toxicology

Date 2022 May 27

PMID 35622602

Authors

Kyle S Gregory

Tejaswini B Mahadeva

Sai Man Liu

K Ravi Acharya

Affiliations

Soon will be listed here.

Abstract

Botulinum neurotoxins (BoNT) are a group of clostridial toxins that cause the potentially fatal neuroparalytic disease botulism. Although highly toxic, BoNTs are utilized as therapeutics to treat a range of neuromuscular conditions. Several serotypes (BoNT/A-/G, /X) have been identified with vastly differing toxicological profiles. Each serotype can be further sub-categorised into subtypes due to subtle variations in their protein sequence. These minor changes have been attributed to differences in both the duration of action and potency for BoNT/A subtypes. BoNTs are composed of three domains-a cell-binding domain, a translocation domain, and a catalytic domain. In this paper, we present the crystal structures of the botulinum neurotoxin A2 cell binding domain, both alone and in complex with its receptor ganglioside GD1a at 1.63 and 2.10 Å, respectively. The analysis of these structures reveals a potential redox-dependent Lys-O-Cys bridge close to the ganglioside binding site and a hinge motion between the H and H subdomains. Furthermore, we make a detailed comparison with the previously reported H/A2:SV2C structure for a comprehensive structural analysis of H/A2 receptor binding.

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References

Zhang S, Masuyer G, Zhang J, Shen Y, Lundin D, Henriksson L . Identification and characterization of a novel botulinum neurotoxin. Nat Commun. 2017; 8:14130. PMC: 5543303. DOI: 10.1038/ncomms14130. View

Yamazaki M, Nelson R, Montal M . Formation of ion channels in lipid bilayers by a peptide with the predicted transmembrane sequence of botulinum neurotoxin A. Protein Sci. 1995; 4(8):1490-7. PMC: 2143195. DOI: 10.1002/pro.5560040806. View

Vagin A, Steiner R, Lebedev A, Potterton L, McNicholas S, Long F . REFMAC5 dictionary: organization of prior chemical knowledge and guidelines for its use. Acta Crystallogr D Biol Crystallogr. 2004; 60(Pt 12 Pt 1):2184-95. DOI: 10.1107/S0907444904023510. View

Hoch D, Ehrlich B, Finkelstein A, Dasgupta B, Simpson L . Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes. Proc Natl Acad Sci U S A. 1985; 82(6):1692-6. PMC: 397338. DOI: 10.1073/pnas.82.6.1692. View

McNicholas S, Potterton E, Wilson K, Noble M . Presenting your structures: the CCP4mg molecular-graphics software. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):386-94. PMC: 3069754. DOI: 10.1107/S0907444911007281. View

Davies J, Hackett G, Liu S, Acharya K . High resolution crystal structures of the receptor-binding domain of neurotoxin serotypes A and FA. PeerJ. 2018; 6:e4552. PMC: 5866713. DOI: 10.7717/peerj.4552. View

Montecucco C, Schiavo G, Rossetto O . The mechanism of action of tetanus and botulinum neurotoxins. Arch Toxicol Suppl. 1996; 18:342-54. DOI: 10.1007/978-3-642-61105-6_32. View

Benoit R, Scharer M, Wieser M, Li X, Frey D, Kammerer R . Crystal structure of the BoNT/A2 receptor-binding domain in complex with the luminal domain of its neuronal receptor SV2C. Sci Rep. 2017; 7:43588. PMC: 5333631. DOI: 10.1038/srep43588. View

Liebschner D, Afonine P, Baker M, Bunkoczi G, Chen V, Croll T . Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr D Struct Biol. 2019; 75(Pt 10):861-877. PMC: 6778852. DOI: 10.1107/S2059798319011471. View

10.

Chen Z, Morris Jr J, Rodriguez R, Wagle Shukla A, Tapia-Nunez J, Okun M . Emerging opportunities for serotypes of botulinum neurotoxins. Toxins (Basel). 2012; 4(11):1196-222. PMC: 3509704. DOI: 10.3390/toxins4111196. View

11.

Gustafsson R, Zhang S, Masuyer G, Dong M, Stenmark P . Crystal Structure of Botulinum Neurotoxin A2 in Complex with the Human Protein Receptor SV2C Reveals Plasticity in Receptor Binding. Toxins (Basel). 2018; 10(4). PMC: 5923319. DOI: 10.3390/toxins10040153. View

12.

McCoy A, Grosse-Kunstleve R, Adams P, Winn M, Storoni L, Read R . Phaser crystallographic software. J Appl Crystallogr. 2009; 40(Pt 4):658-674. PMC: 2483472. DOI: 10.1107/S0021889807021206. View

13.

Davies J, Britton A, Liu S, Acharya K . High-resolution crystal structures of the botulinum neurotoxin binding domains from subtypes A5 and A6. FEBS Open Bio. 2020; 10(8):1474-1481. PMC: 7396429. DOI: 10.1002/2211-5463.12931. View

14.

Williams C, Headd J, Moriarty N, Prisant M, Videau L, Deis L . MolProbity: More and better reference data for improved all-atom structure validation. Protein Sci. 2017; 27(1):293-315. PMC: 5734394. DOI: 10.1002/pro.3330. View

15.

Winn M, Ballard C, Cowtan K, Dodson E, Emsley P, Evans P . Overview of the CCP4 suite and current developments. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):235-42. PMC: 3069738. DOI: 10.1107/S0907444910045749. View

16.

Berman H, Henrick K, Nakamura H . Announcing the worldwide Protein Data Bank. Nat Struct Biol. 2003; 10(12):980. DOI: 10.1038/nsb1203-980. View

17.

Veevers R, Hayward S . Methodological improvements for the analysis of domain movements in large biomolecular complexes. Biophys Physicobiol. 2020; 16:328-336. PMC: 6975908. DOI: 10.2142/biophysico.16.0_328. View

18.

Pellett S, Bradshaw M, Tepp W, Pier C, Whitemarsh R, Chen C . The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes. mBio. 2018; 9(2). PMC: 5874905. DOI: 10.1128/mBio.00089-18. View

19.

Waterman D, Winter G, Gildea R, Parkhurst J, Brewster A, Sauter N . Diffraction-geometry refinement in the DIALS framework. Acta Crystallogr D Struct Biol. 2016; 72(Pt 4):558-75. PMC: 4822564. DOI: 10.1107/S2059798316002187. View

20.

Hill K, Xie G, Foley B, Smith T . Genetic diversity within the botulinum neurotoxin-producing bacteria and their neurotoxins. Toxicon. 2015; 107(Pt A):2-8. DOI: 10.1016/j.toxicon.2015.09.011. View