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Plant ethylene receptors (ETRs) form complicated protein complexes at endoplasmic reticulum membranes. The receptors function as dimers, and they gather to form higher-order protein complexes, together with other proteins that modulate ethylene signaling. Co-immunoprecipitation is a useful biochemical tool for determining the composition of protein complexes. Here, we introduce a basic co-immunoprecipitation scheme that includes preparation of microsomal fraction, immunoprecipitation, and mass spectrometry (MS) analysis for protein identification. Although the ETR protein complex in tomato fruit is addressed here, the protocol may be adapted to other fruit species with some modifications.

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Deciphering Ethylene Signaling via Protein–Protein Interaction in Tomato Fruit

  • 实验方案

  • 发表日期：

    2025年

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  • Yusuke Kamiyoshihara,
  • Yuki Achiha,
  • Shin Ishikawa,
  • Shinji Mizuno,
  • Hitoshi Mori,
  • Akira Tateishi,
  • Donald J. Huber,
  • Harry J. Klee

摘要

Plant ethylene receptors (ETRs) form complicated protein complexes at endoplasmic reticulum membranes. The receptors function as dimers, and they gather to form higher-order protein complexes, together with other proteins that modulate ethylene signaling. Co-immunoprecipitation is a useful biochemical tool for determining the composition of protein complexes. Here, we introduce a basic co-immunoprecipitation scheme that includes preparation of microsomal fraction, immunoprecipitation, and mass spectrometry (MS) analysis for protein identification. Although the ETR protein complex in tomato fruit is addressed here, the protocol may be adapted to other fruit species with some modifications.