Structure-function studies of Borrelia metal transport A (BmtA) from Lyme disease-causing Borrelia burgdorferi

Site-directed mutagenesis and functional analyses in a yeast expression system, combined with PAR metal-binding and ICP-MS assays, identified residues H133, E137, and E203 as essential for Mn²˖ transport. Zinc was also found to stabilize BmtA, with deficiency resulting in protein degradation.<br><br>For biochemical characterization, optimal solubilization and purification were achieved using n-Dodecyl-β-D-Maltopyranoside (DDM), maintaining the protein’s dimeric stat as confirmed by Blue Native PAGE and crosslinking experiments.

Metal-binding and thermal shift assays demonstrated that Mn²˖ induced conformational stabilization, whereas Zn²˖ had minimal impact. Mutations at key residues disrupted folding and stability, highlighting the importance of metal coordination.Overall, this study enhance understanding of BmtA’s structure–function relationship and supports its potential as a therapeutic target for combating Lyme disease.<br><br><i>Thesis is embargoed until 31 December 2026</i>.