Synthesis and Application of a Caged Bioluminescent Probe for the Immunoproteasome DOI
Cody A. Loy, Darci J. Trader

Current Protocols, Journal Year: 2024, Volume and Issue: 4(11)

Published: Nov. 1, 2024

Abstract Monitoring the catalytic activity of proteasome and its various isoforms has become increasingly important with continued development core particle inhibitors targeted protein degraders as potential therapies for diseases high accumulation. The immunoproteasome (iCP) is expressed in a variety due to inflammatory signals, such interferon‐gamma, that alert cell begin generating iCP preferentially over standard proteasome. There need understand expression both cells vivo because it becoming widely isoform diseases. Activity‐based probes have been developed, but their application limited difficult synthesis choice fluorescent reporter. yet be selective probe developed incorporates luminescent reporter could applied different applications. protocols presented here describe cleavable activity‐based bioluminescent iCP, synthesized assays using plate reader. Having this reporter, better understanding how implicated disease progression, well identification small molecule interactors, can achieved. © 2024 Wiley Periodicals LLC. Basic Protocol 1 : Synthesis 2 Expression 3 Immunoproteasome live reader

Language: Английский

Immunoproteasome as a Target for Prodrugs DOI
Christine S. Muli, Cody A. Loy, Darci J. Trader

et al.

Journal of Medicinal Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 17, 2025

Immunoproteasome (iCP) is a proteasome isoform that expressed under inflammatory conditions such as cytokine interferon-γ exposure. The iCP has different catalytic subunits other than the standard CP (standard core particle), allowing production of major histocompatibility complex class I (MHC-I) compatible peptides for eventual T-cell activation. We have previously reported design fluorescent probe monitors activity in cells called TBZ-1, and we applied TBZ-1's recognition sequence prodrug release into iCP-active cells. Here, demonstrate proof-of-concept enzyme. "payload" utilized was toxic moiety, doxorubicin, degrader transcription factor, BRD4. Both examples show required to elicit cell death or degradation This report highlights viable target, its can be used variety cargo expressing iCP.

Language: Английский

Citations

0
Synthesis and Application of a Caged Bioluminescent Probe for the Immunoproteasome DOI
Cody A. Loy, Darci J. Trader

Current Protocols, Journal Year: 2024, Volume and Issue: 4(11)

Published: Nov. 1, 2024

Abstract Monitoring the catalytic activity of proteasome and its various isoforms has become increasingly important with continued development core particle inhibitors targeted protein degraders as potential therapies for diseases high accumulation. The immunoproteasome (iCP) is expressed in a variety due to inflammatory signals, such interferon‐gamma, that alert cell begin generating iCP preferentially over standard proteasome. There need understand expression both cells vivo because it becoming widely isoform diseases. Activity‐based probes have been developed, but their application limited difficult synthesis choice fluorescent reporter. yet be selective probe developed incorporates luminescent reporter could applied different applications. protocols presented here describe cleavable activity‐based bioluminescent iCP, synthesized assays using plate reader. Having this reporter, better understanding how implicated disease progression, well identification small molecule interactors, can achieved. © 2024 Wiley Periodicals LLC. Basic Protocol 1 : Synthesis 2 Expression 3 Immunoproteasome live reader

Language: Английский

Citations

0