Biochemical and Biophysical Investigation of a Calmodulin‐Like Protein From Glycine max Delineates Its Role as a Calcium Sensor During Herbivory

The environment surrounding plants is far from stable, compelling the plant to perceive and adapt to numerous biotic and abiotic constraints, including insect attacks. The perception of a feeding insect typically entails the identification of herbivore‐associated molecular patterns causing a sequential increase in cytosolic Ca2+ levels. Calmodulin‐like proteins (CMLs) are Ca2+ sensor proteins with conserved EF‐hands, which decode Ca2+ signals to generate a stress‐specific response. Although few of the CMLs have been investigated and their role in plant defence has been deciphered in model plants, the role of CMLs as a Ca2+ sensor protein and their interaction mechanism with calcium and downstream targets remains poorly understood in leguminous crop plants. Herein, we demonstrated the presence of a Ca2+ signature in Glycine max during herbivory. Gene expression analysis of GmCML77 (a member of the CML family) indicated its upregulation during S. litura infestation. Phylogenetic analysis and in silico studies predicted GmCML77 as a Ca2+ binding protein. Employing computational modelling and MD simulations, we showed that GmCML77 has predominantly α‐helical conformation with 3 functional Ca2+ binding loops. Also, Ca2+ binding initiates an expansion of tertiary structure, leading to the exposure of hydrophobic residues that may be implicated in its interaction with target proteins. Moreover, gel shift assay and CD spectroscopy results confirmed the Ca2+ binding ability of GmCML77. Our study demonstrated that GmCML77 is a functional calcium‐binding protein, which exhibits conformational changes on Ca2+ binding and acts as a Ca2+ sensor during insect infestation.

• Publication year

  2025

• Venue

  Physiologia Plantarum : An International Journal for Plant Biology

• Publication date

  2025-08-27

• Fields of study

  Biology, Medicine, Chemistry, Environmental Science

• Identifiers
  DOI 10.1111/ppl.70480PMID 40862337
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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