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标题:Biophysical and biological evaluation of optimized stapled peptide inhibitors of the linear ubiquitin chain assembly complex (LUBAC).
时间:2019-05-16 10:33:11
DOI:10.1016/j.bmc.2017.11.047
PMID:29246782
作者:Aguilar-Alonso F;Whiting AL;Kim YJ;Bernal F
出版源: 《Bioorg Med Chem》 ,2018 ,26 (6) :S0968089617317467
摘要:Biophysical and biological evaluation of optimized stapled peptide inhibitors of the linear ubiquitin chain assembly complex (LUBAC)Abstract Linear ubiquitylation, in which ubiquitin units are covalently linked through N- and C-terminal amino ...
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页数:11 PAGES
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目录:
  • Biophysical and biological evaluation of optimized stapled peptide inhibitors of the linear ubiquitin chain assembly complex (LUBAC)
    • 1 Introduction
    • 2 Results and discussion
      • 2.1 Design and synthesis of stapled HOIP peptides
      • 2.2 Stapled HOIP peptides inhibit LUBAC ubiquitylation activity in␣vitro
      • 2.3 Stapled HOIP peptides unexpectedly bind to HOIP-UBA over HOIL-UBL
      • 2.4 Active HOIP peptides affect cancer cell viability through the NF-κB pathway
      • 2.5 Active HOIP peptides exhibit inhibitory effects on the NF-κB pathway following TNFα stimulation
    • 3 Conclusions
    • 4 Experimental section
      • 4.1 Chemistry procedures
        • 4.1.1 Peptide synthesis and purification
        • 4.1.2 Circular dichroism spectroscopy
        • 4.1.3 Surface plasmon resonance (SPR)
      • 4.2 Cell lines and maintenance
      • 4.3 Preparation of purified proteins
      • 4.4 In vitro autoubiquitylation assays
      • 4.5 High resolution clear native electrophoresis (hrCNE)
      • 4.6 Cell viability assay
      • 4.7 Cell cytotoxicity and apoptosis assay
      • 4.8 Western blots
      • 4.9 Peptide cell permeability
      • 4.10 TNF-α stimulation, NF-κB p65 DNA binding, and IL-8 ELISAs
      • 4.11 Quantification and statistical analysis
    • Acknowledgments
    • ack26
    • Funding
    • A Supplementary data
    • References

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