At1g79710 Antibody

Molecular Characterization of AT1G79710

Protein Structure:

• AT1G79710 encodes a 510-amino-acid protein with 12 predicted transmembrane domains, characteristic of MFS transporters .

• Theoretical molecular weight: ~56 kDa .

Functional Domains:

The antibody typically targets epitopes in the cytoplasmic N-terminal region (residues 1–44), which are surface-exposed and less hydrophobic .

Development and Validation of the Antibody

Immunogen Design:

• A synthetic peptide corresponding to residues 12–31 (EDLKRQISNSSVKETLQKRL) was used for immunization in rabbits .

• This region was selected due to high antigenicity scores and low homology to other MFS proteins .

Validation Data:

Cross-reactivity tests confirmed no binding to related MFS proteins (e.g., AT1G79720 or AT2G32040) .

3.1. Functional Studies

• Subcellular Localization: The antibody confirmed AT1G79710's plasma membrane localization in root epidermal cells, consistent with its role in nutrient uptake .

• Knockdown Phenotypes:

  • AT1G79710-silenced plants showed 40% reduced growth under low-phosphate conditions ($p < 0.01$) .

  • Altered metabolite profiles (e.g., 2.5-fold increase in cytosolic sucrose) .

3.2. Protein-Protein Interactions

Co-immunoprecipitation studies identified interactions with:

• AT5G54860 (ABC transporter)

• AT1G31300 (lipid-sensing protein)
  These interactions suggest a role in coordinating nutrient and lipid transport .

Challenges in Antibody Specificity

Despite rigorous validation, limitations include:

• Sample Preparation: Requires nonionic detergents (e.g., 0.1% Triton X-100) to prevent epitope masking .

• Tissue Specificity: Strongest signal in roots; weaker expression in leaves .

• Structural Biology: Cryo-EM studies using the antibody could resolve transporter conformations .

• Biosensor Development: Engineered nanobodies derived from AT1G79710 antibodies may enable real-time nutrient monitoring .