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Interaction of Synthetic Human SLURP-1 with the Nicotinic Acetylcholine Receptors

Journal Article


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Abstract


  • Human SLURP-1 is a secreted protein of the Ly6/uPAR/three-finger neurotoxin family that co-localizes with nicotinic acetylcholine receptors (nAChRs) and modulates their functions. Conflicting biological activities of SLURP-1 at various nAChR subtypes have been based on heterologously produced SLURP-1 containing N- and/or C-terminal extensions. Here, we report the chemical synthesis of the 81 amino acid residue human SLURP-1 protein, characterization of its 3D structure by NMR, and its biological activity at nAChR subtypes. Radioligand assays indicated that synthetic SLURP-1 did not compete with [125I]-α-bungarotoxin (α-Bgt) binding to human neuronal α7 and Torpedo californica muscle-type nAChRs, nor to mollusk acetylcholine binding proteins (AChBP). Inhibition of human α7-mediated currents only occurred in the presence of the allosteric modulator PNU120596. In contrast, we observed robust SLURP-1 mediated inhibition of human α3β4, α4β4, α3β2 nAChRs, as well as human and rat α9α10 nAChRs. SLURP-1 inhibition of α9α10 nAChRs was accentuated at higher ACh concentrations, indicating an allosteric binding mechanism. Our results are discussed in the context of recent studies on heterologously produced SLURP-1 and indicate that N-terminal extensions of SLURP-1 may affect its activity and selectivity on its targets. In this respect, synthetic SLURP-1 appears to be a better probe for structure-function studies.

Authors


  •   Durek, Thomas (external author)
  •   Shelukhina, Irina V. (external author)
  •   Tae, Han Shen
  •   Thongyoo, Panumart (external author)
  •   Spirova, Ekaterina N. (external author)
  •   Kudryavtsev, Denis S. (external author)
  •   Kasheverov, Igor E. (external author)
  •   Faure, Grazyna (external author)
  •   Corringer, Pierre-Jean (external author)
  •   Craik, David J. (external author)
  •   Adams, David J.
  •   Tsetlin, Victor I. (external author)

Publication Date


  • 2017

Citation


  • Durek, T., Shelukhina, I. V., Tae, H., Thongyoo, P., Spirova, E. N., Kudryavtsev, D. S., Kasheverov, I. E., Faure, G., Corringer, P., Craik, D. J., Adams, D. J. & Tsetlin, V. I. (2017). Interaction of Synthetic Human SLURP-1 with the Nicotinic Acetylcholine Receptors. Scientific Reports, 7 16606-1-16606-11.

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=2218&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1191

Start Page


  • 16606-1

End Page


  • 16606-11

Volume


  • 7

Place Of Publication


  • United Kingdom

Abstract


  • Human SLURP-1 is a secreted protein of the Ly6/uPAR/three-finger neurotoxin family that co-localizes with nicotinic acetylcholine receptors (nAChRs) and modulates their functions. Conflicting biological activities of SLURP-1 at various nAChR subtypes have been based on heterologously produced SLURP-1 containing N- and/or C-terminal extensions. Here, we report the chemical synthesis of the 81 amino acid residue human SLURP-1 protein, characterization of its 3D structure by NMR, and its biological activity at nAChR subtypes. Radioligand assays indicated that synthetic SLURP-1 did not compete with [125I]-α-bungarotoxin (α-Bgt) binding to human neuronal α7 and Torpedo californica muscle-type nAChRs, nor to mollusk acetylcholine binding proteins (AChBP). Inhibition of human α7-mediated currents only occurred in the presence of the allosteric modulator PNU120596. In contrast, we observed robust SLURP-1 mediated inhibition of human α3β4, α4β4, α3β2 nAChRs, as well as human and rat α9α10 nAChRs. SLURP-1 inhibition of α9α10 nAChRs was accentuated at higher ACh concentrations, indicating an allosteric binding mechanism. Our results are discussed in the context of recent studies on heterologously produced SLURP-1 and indicate that N-terminal extensions of SLURP-1 may affect its activity and selectivity on its targets. In this respect, synthetic SLURP-1 appears to be a better probe for structure-function studies.

Authors


  •   Durek, Thomas (external author)
  •   Shelukhina, Irina V. (external author)
  •   Tae, Han Shen
  •   Thongyoo, Panumart (external author)
  •   Spirova, Ekaterina N. (external author)
  •   Kudryavtsev, Denis S. (external author)
  •   Kasheverov, Igor E. (external author)
  •   Faure, Grazyna (external author)
  •   Corringer, Pierre-Jean (external author)
  •   Craik, David J. (external author)
  •   Adams, David J.
  •   Tsetlin, Victor I. (external author)

Publication Date


  • 2017

Citation


  • Durek, T., Shelukhina, I. V., Tae, H., Thongyoo, P., Spirova, E. N., Kudryavtsev, D. S., Kasheverov, I. E., Faure, G., Corringer, P., Craik, D. J., Adams, D. J. & Tsetlin, V. I. (2017). Interaction of Synthetic Human SLURP-1 with the Nicotinic Acetylcholine Receptors. Scientific Reports, 7 16606-1-16606-11.

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=2218&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1191

Start Page


  • 16606-1

End Page


  • 16606-11

Volume


  • 7

Place Of Publication


  • United Kingdom