Single-domain antibodies for functional targeting of the signaling scaffold Shoc2

Hye In Jang, Patricia G. Wilson, Mary Sau, Udeep Chawla, David W. Rodgers, Emilia Galperin

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The accurate transmission of signals by the canonical ERK1/2 kinase pathway critically relies on the proper assembly of an intricate multiprotein complex by the scaffold protein Shoc2. However, the details of the mechanism by which Shoc2 guides ERK1/2 signals are not clear, in part, due to the lack of research tools targeting specific protein binding moieties of Shoc2. We report generation and characterization of single domain antibodies against human Shoc2 using a universal synthetic library of humanized nanobodies. Our results identify eight synthetic single-domain antibodies and show that two evaluated antibodies have binding affinities to Shoc2 in the nanomolar range. High affinity antibodies were uniquely suited for the analysis of the Shoc2 complex assembly. Selected single-domain antibodies were also functional in intracellular assays. This study illustrates that Shoc2 single-domain antibodies can be used to understand functional mechanisms governing complex multiprotein signaling modules and have promise in application for therapies that require modulation of the ERK1/2-associated diseases.

Original languageEnglish
Pages (from-to)110-116
Number of pages7
JournalMolecular Immunology
Volume118
DOIs
StatePublished - Feb 2020

Bibliographical note

Funding Information:
We thank Dr. Craig Vander Kooi for critical reading of the manuscript and Dr. Tianyan Gao for sharing valuable reagents. This project was supported by grants from the National Institute of General Medical Sciences ( GM113087 to EG) and the American Cancer Society ( RSG-14-172-01-CSM to EG). Its content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Health.

Publisher Copyright:
© 2019 Elsevier Ltd

Keywords

  • ERK1/2 signaling
  • Scaffold
  • Shoc2
  • Single domain antibody

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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