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Updated: Aug 20, 2025

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
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Differential dynamics of cullin deneddylation via COP9 signalosome subunit 5 interaction

Yeong-Mu Kim1, Hye-Ji Kim1, Dong-Kyu Kim1

  • 1Department of Biochemistry, Chungbuk National University, Cheongju, 28644, Republic of Korea.

Biochemical and Biophysical Research Communications
|November 24, 2022
PubMed

Insights

Cullin-RING ligase (CRL) deneddylation rates vary, with CUL2 showing slower dynamics due to weaker binding to CSN5. This differential regulation impacts CRL function in protein degradation.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Cullin-RING ligases (CRLs) are crucial for proteasomal degradation, regulating key cellular processes like cell cycle control and genome stability.
  • CRL activity is modulated by neddylation (activation) and deneddylation (inactivation) mediated by the COP9 signalosome (CSN).

Purpose of the Study:

  • To investigate the differential deneddylation rates among various Cullin (CUL) proteins.
  • To elucidate the underlying mechanisms governing the distinct neddylation-deneddylation dynamics of CULs.

Main Methods:

  • Treatment with pevonedistat, a NEDD8-activating enzyme (NAE) inhibitor.
  • Immunoprecipitation assays to assess binding affinities between CULs and CSN5.
  • Analysis of deneddylation kinetics following CSN5 inhibition.

Main Results:

  • Pevonedistat treatment rapidly inhibited neddylation in CUL1, CUL3, CUL4A/B, and CUL5, but slowly increased CUL2 deneddylation.
  • CUL2 exhibits a lower binding affinity to CSN5 compared to other CULs.
  • CSN5 inhibition resulted in slower deneddylation of CUL2 relative to other CULs.

Conclusions:

  • The binding strength between CULs and CSN5 dictates their deneddylation rates.
  • CUL2's distinct deneddylation dynamics are attributed to its weaker interaction with CSN5.
  • These findings offer mechanistic insights into the differential regulation of CRLs.

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