Plate-based assay for measuring direct semaphorin-neuropilin interactions

Matthew W. Parker, Craig W. Vander Kooi

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The semaphorins are an essential family of axon guidance molecules that can be either secreted or are transmembrane proteins. Class 3 semaphorin (Sema3) family members are secreted and provide longrange guidance cues through two receptor families: neuropilins (Nrp) and plexins. Nrp is uniquely required for high-affinity Sema3 binding and signaling. Therefore, characterizing the molecular details of the Sema3/Nrp interaction is important for understanding the broader physiological and pathological role of the Sema3 family of proteins. Here we describe an in vitro plate-based binding assay for characterization of the Sema3/Nrp interaction. This assay utilizes Nrp-affinity plates and an alkaline phosphatase (AP)-Sema3 fusion to rapidly measure direct Sema3/Nrp binding. This assay can be used to measure receptorligand binding, the contribution of different domains, and exogenous factors, and to characterize competitive ligand binding.

Original languageEnglish (US)
Pages (from-to)73-87
Number of pages15
JournalMethods in Molecular Biology
Volume1493
DOIs
StatePublished - 2017
Externally publishedYes

Keywords

  • Assay
  • Binding
  • Competition
  • Interaction
  • Ligand
  • Neuropilin
  • Plate-based
  • Receptor
  • Semaphorin

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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