Covalently Engineered Nanobody Chimeras for Targeted Membrane Protein Degradation

J Am Chem Soc. 2021 Oct 13;143(40):16377-16382. doi: 10.1021/jacs.1c08521. Epub 2021 Oct 1.

Abstract

The targeted degradation of membrane proteins would afford an attractive and general strategy for treating various diseases that remain difficult with the current proteolysis-targeting chimera (PROTAC) methodology. We herein report a covalent nanobody-based PROTAC strategy, termed GlueTAC, for targeted membrane protein degradation with high specificity and efficiency. We first established a mass-spectrometry-based screening platform for the rapid development of a covalent nanobody (GlueBody) that allowed proximity-enabled cross-linking with surface antigens on cancer cells. By conjugation with a cell-penetrating peptide and a lysosomal-sorting sequence, the resulting GlueTAC chimera triggered the internalization and degradation of programmed death-ligand 1 (PD-L1), which provides a new avenue to target and degrade cell-surface proteins.

MeSH terms

  • B7-H1 Antigen / chemistry
  • B7-H1 Antigen / metabolism
  • Cell Line, Tumor
  • Cell-Penetrating Peptides / chemistry
  • Cell-Penetrating Peptides / metabolism
  • Humans
  • Membrane Proteins* / chemistry
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Protein Engineering
  • Proteolysis*
  • Single-Domain Antibodies* / chemistry

Substances

  • Single-Domain Antibodies
  • Membrane Proteins
  • B7-H1 Antigen
  • Cell-Penetrating Peptides