Figure 4

Identification of M . loti Ca ²⁺ binding proteins by ⁴⁵ Ca ²⁺ -ligand overlay. Proteins were dot-blotted onto nitrocellulose and then incubated with ⁴⁵CaCl₂ in a calcium overlay assay [44]. ⁴⁵Ca-labeled proteins were visualized by autoradiography (7-d exposure). 1, equine cytochrome c, used as negative control (5 μg); 2, purified M. loti 20 kDa Stains-all blue-staining protein (5 μg); 3, rabbit calsequestrin, used as positive control (1 μg).