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TitleStructure and analysis of nanobody binding to the human ASIC1a ion channel.
Journal, issue, pagesElife, Vol. 10, Year 2021
Publish dateJul 28, 2021
AuthorsYangyu Wu / Zhuyuan Chen / Fred J Sigworth / Cecilia M Canessa /
PubMed AbstractASIC1a is a proton-gated sodium channel involved in modulation of pain, fear, addiction, and ischemia-induced neuronal injury. We report isolation and characterization of alpaca-derived nanobodies ...ASIC1a is a proton-gated sodium channel involved in modulation of pain, fear, addiction, and ischemia-induced neuronal injury. We report isolation and characterization of alpaca-derived nanobodies (Nbs) that specifically target human ASIC1a. Cryo-electron microscopy of the human ASIC1a channel at pH 7.4 in complex with one of these, Nb.C1, yielded a structure at 2.9 Å resolution. It is revealed that Nb.C1 binds to a site overlapping with that of the Texas coral snake toxin (MitTx1) and the black mamba venom Mambalgin-1; however, the Nb.C1-binding site does not overlap with that of the inhibitory tarantula toxin psalmotoxin-1 (PcTx1). Fusion of Nb.C1 with PcTx1 in a single polypeptide markedly enhances the potency of PcTx1, whereas competition of Nb.C1 and MitTx1 for binding reduces channel activation by the toxin. Thus, Nb.C1 is a molecular tool for biochemical and structural studies of hASIC1a; a potential antidote to the pain-inducing component of coral snake bite; and a candidate to potentiate PcTx1-mediated inhibition of hASIC1a in vivo for therapeutic applications.
External linksElife / PubMed:34319232 / PubMed Central
MethodsEM (single particle)
Resolution2.86 Å
Structure data

24580

7rnn

EMDB-24580, PDB-7rnn:
Human ASIC1a-Nb.C1 complex
Method: EM (single particle) / Resolution: 2.86 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

Source
  • homo sapiens (human)
  • vicugna pacos (alpaca)
KeywordsMEMBRANE PROTEIN / Channel Proton-gated Nanobody

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