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Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors
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The chloromethylketone protease inhibitor AAPF(CMK) also targets ATP-dependent helicases and SAP-domain proteins

Chetan Dhamne1, David A Drubin, Kimberly Duncan

  • 1Gittlen Cancer Research Foundation, Pennsylvania State University, Hershey, PA 17033, USA.

Journal of Cellular Biochemistry
|September 29, 2006
PubMed

Insights

The inhibitor AAPF(CMK) targets nuclear proteases and affects cellular transformation. It also inhibits ATP-dependent helicases, including SV40 large T antigen, impacting DNA repair and chromatin remodeling.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Virology

Background:

  • A nuclear protease implicated in cellular transformation and high-risk human papillomavirus (HPV) infections has been under investigation.
  • This protease exhibits chymotrypsin-like substrate specificity and is potently inhibited by AAPF(CMK).
  • Recent observations suggest AAPF(CMK) exerts effects independent of nuclear protease inhibition in certain model systems.

Purpose of the Study:

  • To elucidate the broader reactivity and inhibitory targets of AAPF(CMK) beyond the nuclear protease.
  • To investigate the interaction of AAPF(CMK) with ATP-dependent helicases and other nuclear complexes.
  • To determine if AAPF(CMK) affects the helicase activity of specific proteins like SV40 large T antigen.

Main Methods:

  • Chemical inhibition assays using AAPF(CMK).
  • Proteomic analysis to identify proteins that selectively react with AAPF(CMK).
  • In vitro assays to assess the effect of AAPF(CMK) on protein helicase activity.

Main Results:

  • AAPF(CMK) selectively reacts with ATP-dependent helicases.
  • The inhibitor also reacts with a limited range of proteins within DNA repair/chromatin remodeling complexes, such as Cohesin components and SAP-domain proteins.
  • In vitro, AAPF(CMK) specifically reacts with SV40 large T antigen and inhibits its helicase activity.

Conclusions:

  • The chloromethylketone inhibitor AAPF(CMK) possesses broader inhibitory activity than previously recognized, targeting ATP-dependent helicases.
  • These findings reveal novel interactions of AAPF(CMK) with key nuclear complexes involved in DNA repair and chromatin remodeling.
  • AAPF(CMK) serves as a valuable tool for studying the function of helicases, including viral proteins like SV40 large T antigen.

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