Human S100A5 binds Ca and Cu independently.

Background: S100A5 is a calcium binding protein found in a small subset of amniote tissues. Little is known about the biological roles of S100A5, but it may be involved in inflammation and olfactory signaling. Previous work indicated that S100A5 displays antagonism between binding of Ca and Cu ions-one of the most commonly cited features of the protein. We set out to characterize the interplay between Ca and Cu binding by S100A5 using isothermal titration calorimetry (ITC), circular dichroism spectroscopy (CD), and analytical ultracentrifugation (AUC).

Results: We found that human S100A5 is capable of binding both Cu and Ca ions simultaneously. The wildtype protein was extremely aggregation-prone in the presence of Cu and Ca. A Cys-free version of S100A5, however, was not prone to precipitation or oligomerization. Mutation of the cysteines does not disrupt the binding of either Ca or Cu to S100A5. In the Cys-free background, we measured Ca and Cu binding in the presence and absence of the other metal using ITC. Saturating concentrations of Ca or Cu do not disrupt the binding of one another. Ca and Cu binding induce structural changes in S100A5, which are measurable using CD spectroscopy. We show via sedimentation velocity AUC that the wildtype protein is prone to the formation of soluble oligomers, which are not present in Cys-free samples.

Conclusions: S100A5 can bind Ca and Cu ions simultaneously and independently. This observation is in direct contrast to previously-reported antagonism between binding of Cu and Ca ions. The previous result is likely due to metal-dependent aggregation. Little is known about the biology of S100A5, so an accurate understanding of the biochemistry is necessary to make informed biological hypotheses. Our observations suggest the possibility of independent biological functions for Cu and Ca binding by S100A5.