Chemical and Enzymatic Approaches to C-Terminal Cyclic Imide Formation.

Zhenguang Zhao, Ethan Yang Feng, Wenqing Xu, Christina M Woo

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Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

4 authors.

Zhenguang Zhao *Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.

Ethan Yang Feng *Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.

Wenqing Xu *Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.

Christina M WooDepartment of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA. cwoo@chemistry.harvard.edu.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The C-terminal cyclic imide modification is recognized as a degron by the E3 ligase adapter cereblon (CRBN). Reliable methods to generate and measure this modification on CRBN substrates are essential for studying its biological function. In this chapter, we describe two complementary approaches for producing C-terminal cyclic imide-modified peptides and proteins: sortase-mediated transpeptidation and enzymatic conversion by protein carboxymethyltransferase (PCMT1). Additionally, we present analytical strategies, including a time-resolved Förster resonance energy transfer (TR-FRET) assay for measuring binding of substrates to CRBN in vitro and mass spectrometry methods for confirming cyclic imide formation. Together, these protocols provide practical methods for investigating the chemistry and biology of C-terminal cyclic imides.

Indexed as

ImidesUbiquitin-Protein LigasesAdaptor Proteins, Signal TransducingAminoacyltransferasesCysteine EndopeptidasesDegronsFluorescence Resonance Energy TransferHumansMass SpectrometryAdaptor Proteins, Signal TransducingAminoacyltransferasesCRBN protein, humanCysteine EndopeptidasesImidesUbiquitin-Protein LigasesCereblonC-terminal cyclic imideMass spectrometryPCMT1SortaseTargeted protein degradationTR-FRET

Identifiers

PMID42573690

DOI10.1038/s41589-021-00877-5

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.