MTP6 Antibody

Antigen and Antibody Development

The MTP6 antibody targets the C-terminal region of CsMTP6, a cucumber (Cucumis sativus) homolog of plant mitochondrial metal transporters. Key aspects include:

• Antigen Design: A synthetic peptide corresponding to residues 324–337 (C+CEGVKGCHRLRGRRA) of CsMTP6 was conjugated to keyhole limpet hemocyanin (KLH) for immunization .

• Host Species: Rabbits were immunized, and IgG was purified using antigen-linked Sepharose resin .

• Validation: Specificity was confirmed via ELISA and western blotting, showing a single band at ~35 kDa in mitochondrial fractions .

Antibody Applications

The MTP6 antibody has been utilized in diverse experimental contexts:

Role in Mitochondrial Metal Transport

• Fe and Mn Homeostasis: CsMTP6 facilitates Fe export from mitochondria to the cytosol, as shown by restored growth in Fe-deficient yeast (Δmmt1Δmmt2) expressing CsMTP6 .

• Mechanistic Insights:

  • Overexpression in yeast increased cytosolic Fe (via c-GDO activity assays) and decreased mitochondrial Fe .

  • Structural analysis revealed conserved metal-binding motifs (HSVSD and HHRAD) critical for transport .

Subcellular Localization

• Mitochondrial Targeting: Immunoblotting and GFP fusion confirmed CsMTP6 localization to mitochondria, with a processed molecular weight of 35 kDa (vs. predicted 50 kDa) .

Validation and Controls

• Knockout Validation: No cross-reactivity observed in non-mitochondrial fractions (plasma membrane, tonoplast, plastid) .

• Functional Complementation: Rescue of yeast Δmmt1Δmmt2 phenotypes confirmed antibody specificity and target functionality .