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Purpose <p>Phosphoinositide-specific phospholipase C plays a crucial role in the trans-membrane and lipid signal transduction. Previous studies have elucidated the biological functions of PI-PLC. However, its function mechanism has not been fully understood.</p> Results <p>We present a novel approach to analyze the in vitro activity of phospholipase C from <i>Populus simonii</i> x <i>P. nigra</i>. The results indicate that the optimal condition for PsnPLC activity is pH 7.4/10&#xa0;μM Ca<sup>2+</sup>. The increase of Ca<sup>2+</sup> and pH, as well as the deletion of the EF-hand domain or C2 domain, will lead to the reduced PsnPLC activity. Electrophoresis migration shift assays confirm that PsnPLC could bind Ca<sup>2+</sup> in vitro, while conditions of Ca<sup>2+</sup> binding and pH 8.8 might lead to its degradation.</p> Conclusion <p>The findings identify Ca<sup>2+</sup> and pH as key factors modulating PsnPLC function, which will provide evidence linking PI-PLC and the Ca<sup>2+</sup> signaling network.</p>

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A novel approach for the in vitro assay of phospholipase C and its application in analyzing the effect of calcium and pH on populus PsnPLC

  • 期刊论文

  • 发表日期：

    2025年9月23日

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  • Yao Sun,
  • Chunhui Yang,
  • Xin Sun,
  • Yao Li,
  • Qiong Wu,
  • Di Fu,
  • Lei Wang

摘要

Purpose

Phosphoinositide-specific phospholipase C plays a crucial role in the trans-membrane and lipid signal transduction. Previous studies have elucidated the biological functions of PI-PLC. However, its function mechanism has not been fully understood.

Results

We present a novel approach to analyze the in vitro activity of phospholipase C from Populus simonii x P. nigra. The results indicate that the optimal condition for PsnPLC activity is pH 7.4/10 μM Ca2+. The increase of Ca2+ and pH, as well as the deletion of the EF-hand domain or C2 domain, will lead to the reduced PsnPLC activity. Electrophoresis migration shift assays confirm that PsnPLC could bind Ca2+ in vitro, while conditions of Ca2+ binding and pH 8.8 might lead to its degradation.

Conclusion

The findings identify Ca2+ and pH as key factors modulating PsnPLC function, which will provide evidence linking PI-PLC and the Ca2+ signaling network.