Quantification of Ca(2+) binding to melanin supports the hypothesis that melanosomes serve a functional role in regulating calcium homeostasis.

Calcium regulation in melanocytes affects numerous biological pathways including protecting the redox balance in the cell and regulating the supply of substrate, l-tyrosine, for melanogenesis. The pigment contained in the melanocytes, melanin, has been implicated in maintaining calcium homeostasis in the cell and is known to be involved with calcium ion regulation in the inner ear. Herein, the association constant for Ca(2+) binding to Sepia melanin is determined by isothermal titration calorimetry to be 3.3 (+/-0.2) x 10(3)/M. This value is comparable with other well-established intracellular calcium-binding proteins that serve to buffer calcium concentrations, lending further support to the hypothesis that melanosomes serve as intracellular mediators of calcium homeostasis in melanocytes. Using this binding constant and the data from a fluorescent Ca(2+) displacement assay, the pK(a) of the carboxyl group coordinated to Ca(2+) is determined to be 3.1 +/- 0.1.