N-linked glycosylation of SV2 is required for binding and uptake of botulinum neurotoxin A.
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.
Department of Physiology and Biophysics, University of California, Irvine, Irvine, California, USA.
Guorui
G
Yao
Department of Urology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.|Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.|Department of Surgery, Harvard Medical School, Boston, Massachusetts, USA.
Sicai
S
Zhang
Institut für Toxikologie, Medizinische Hochschule Hannover, Hannover, Germany.
Stefan
S
Mahrhold
Department of Physiology and Biophysics, University of California, Irvine, Irvine, California, USA.
Kwok-Ho
KH
Lam
Centre for Biological Threats and Special Pathogens-Biological Toxins (ZBS3), Robert Koch-Institut, Berlin, Germany.
Daniel
0000-0001-9057-4283
D
Stern
Department of Molecular Immunology, Beckman Research Institute of City of Hope, Duarte, California, USA.
Karine
K
Bagramyan
NE-CAT, Argonne National Laboratory, Department of Chemistry and Chemical Biology, Cornell University, Argonne, Illinois, USA.
Kay
0000-0002-4046-1704
K
Perry
Department of Molecular Immunology, Beckman Research Institute of City of Hope, Duarte, California, USA.
Markus
M
Kalkum
Institut für Toxikologie, Medizinische Hochschule Hannover, Hannover, Germany.
Andreas
A
Rummel
Department of Urology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.|Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.|Department of Surgery, Harvard Medical School, Boston, Massachusetts, USA.
Min
M
Dong
Department of Physiology and Biophysics, University of California, Irvine, Irvine, California, USA.
Rongsheng
0000-0003-0348-7363
R
Jin
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000931:Antidotes / Q000737:chemistry*; D001665:Binding Sites; D001692:Biological Transport; D019274:Botulinum Toxins, Type A / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D003014:Clostridium botulinum / Q000737:chemistry* / Q000472:pathogenicity; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000931:Antidotes; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; C585899:SV2C protein, human; D019274:Botulinum Toxins, Type A