N-linked glycosylation of SV2 is required for binding and uptake of botulinum neurotoxin A

Guorui Yao; Sicai Zhang; Stefan Mahrhold; Kwok-Ho Lam; Daniel Stern; Karine Bagramyan; Kay Perry; Markus Kalkum; Andreas Rummel; Min Dong; Rongsheng Jin

doi:10.1038/nsmb.3245

N-linked glycosylation of SV2 is required for binding and uptake of botulinum neurotoxin A

Nat Struct Mol Biol. 2016 Jul;23(7):656-62. doi: 10.1038/nsmb.3245. Epub 2016 Jun 13.

Authors

Guorui Yao¹, Sicai Zhang^{2

3

4}, Stefan Mahrhold⁵, Kwok-Ho Lam¹, Daniel Stern⁶, Karine Bagramyan⁷, Kay Perry⁸, Markus Kalkum⁷, Andreas Rummel⁵, Min Dong^{2

3

4}, Rongsheng Jin¹

Affiliations

¹ Department of Physiology and Biophysics, University of California, Irvine, Irvine, California, USA.
² Department of Urology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
³ Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
⁴ Department of Surgery, Harvard Medical School, Boston, Massachusetts, USA.
⁵ Institut für Toxikologie, Medizinische Hochschule Hannover, Hannover, Germany.
⁶ Centre for Biological Threats and Special Pathogens-Biological Toxins (ZBS3), Robert Koch-Institut, Berlin, Germany.
⁷ Department of Molecular Immunology, Beckman Research Institute of City of Hope, Duarte, California, USA.
⁸ NE-CAT, Argonne National Laboratory, Department of Chemistry and Chemical Biology, Cornell University, Argonne, Illinois, USA.

Abstract

Botulinum neurotoxin serotype A1 (BoNT/A1), a licensed drug widely used for medical and cosmetic applications, exerts its action by invading motoneurons. Here we report a 2.0-Å-resolution crystal structure of the BoNT/A1 receptor-binding domain in complex with its neuronal receptor, glycosylated human SV2C. We found that the neuronal tropism of BoNT/A1 requires recognition of both the peptide moiety and an N-linked glycan on SV2. This N-glycan-which is conserved in all SV2 isoforms across vertebrates-is essential for BoNT/A1 binding to neurons and for its potent neurotoxicity. The glycan-binding interface on SV2 is targeted by a human BoNT/A1-neutralizing antibody currently licensed as an antibotulism drug. Our studies reveal a new paradigm of host-pathogen interactions, in which pathogens exploit conserved host post-translational modifications, thereby achieving highly specific receptor binding while also tolerating genetic changes across multiple isoforms of receptors.

MeSH terms

Amino Acid Sequence
Antibodies, Monoclonal / chemistry*
Antidotes / chemistry*
Binding Sites
Biological Transport
Botulinum Toxins, Type A / chemistry*
Botulinum Toxins, Type A / metabolism
Cloning, Molecular
Clostridium botulinum / chemistry*
Clostridium botulinum / pathogenicity
Crystallography, X-Ray
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Glycosylation
HEK293 Cells
Humans
Membrane Glycoproteins / antagonists & inhibitors
Membrane Glycoproteins / chemistry
Membrane Glycoproteins / genetics
Membrane Glycoproteins / metabolism*
Models, Molecular
Nerve Tissue Proteins / antagonists & inhibitors
Nerve Tissue Proteins / chemistry
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism*
Protein Binding
Protein Domains
Protein Processing, Post-Translational*
Protein Structure, Secondary
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism

Substances

Antibodies, Monoclonal
Antidotes
Membrane Glycoproteins
Nerve Tissue Proteins
Recombinant Fusion Proteins
SV2C protein, human
Botulinum Toxins, Type A

Abstract

MeSH terms

Substances

Grants and funding