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QUICK identification and SPR validation of signal transducers and activators of transcription 3 (Stat3) interacting

Peng Zheng1, Qiu Zhong, Qian Xiong

  • 1Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.

Journal of Proteomics
|November 15, 2011
PubMed

Insights

Researchers identified 38 proteins interacting with Signal transducer and activator of transcription 3 (Stat3) in multiple myeloma cells. This study provides new insights into Stat3 regulation and function in human diseases.

Area of Science:

  • Molecular Biology
  • Proteomics
  • Cancer Biology

Background:

  • Signal transducer and activator of transcription 3 (Stat3) plays a role in human disease pathogenesis.
  • Constitutive Stat3 activity is observed in human multiple myeloma (MM) U266 cells.
  • Stat3-regulated mechanisms in MM are not fully understood.

Purpose of the Study:

  • To systematically identify Stat3 interacting proteins in U266 MM cells.
  • To elucidate the regulatory mechanisms of Stat3 activity and function in MM.

Main Methods:

  • Quantitative immunoprecipitation combined with knockdown (QUICK) was employed.
  • QUICK integrates RNAi, stable isotope labeling with amino acids in cell culture (SILAC), immunoprecipitation, and quantitative mass spectrometry (MS).
  • Reciprocal co-immunoprecipitations and surface plasmon resonance (SPR) were used for validation.

Main Results:

  • A total of 38 Stat3 interacting proteins were identified.
  • Proteomic analysis distinguished specific Stat3 interactors from background proteins.
  • 14-3-3ζ, PRKCB, and Hsp90 were confirmed as Stat3 interacting proteins.

Conclusions:

  • The study identified novel Stat3 interacting proteins involved in various cellular functions.
  • New insights into the regulation of Stat3 activity and function in multiple myeloma cells were provided.

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