The evolution of S100A7: an unusual gene expansion in Myotis bats.

BACKGROUND: The S100A7 gene, also called psoriasin, was first described as an upregulated protein in psoriatic skin. For the past years, the importance of this protein as a key effector of innate immunity has been clearly established, not only due to its importance protecting against bacteria skin insult in humans, but also because of its important role in amplifying inflammatory processes. Given the importance of S100A7 in host defense, S100A7 genes have been mostly studied in humans. Here we provide a detailed analysis of the evolution of the gene family encoding for the S100A7 protein in mammals. RESULTS: Examination of several mammalian genomes revealed an unexpected variation in the copy number of S100A7. Among the most representative mammalian groups, we report that multiple events of duplication, gene loss and high mutation rates are shaping the evolution of this gene family. An unexpected result comes from Myotis species (order Chiroptera), where we found an outstanding S100A7 gene radiation, resulting in more than 10 copies in M. lucifugus and 5 copies in M. brandtii. These findings suggest a unique adaptive road in these species and are suggestive of special role of this protein in their immune system. CONCLUSIONS: We found different evolutionary histories among different mammalian groups. Overall, our results suggest that this gene family is evolving under the birth-and-death model of evolution. To our knowledge, this work represents the first detailed analysis of phylogenetic relationships of S100A7 within mammals and therefore will pave the way to further clarify their unique function in the immune system.

Preparation of the Oxidized and Reduced Forms of Psoriasin (S100A7).

Human S100A7 (psoriasin) is a metal-chelating host-defense protein expressed by epithelial cells. S100A7 possesses two Cys residues that generate two redox isoforms of the protein. In the oxidized form (S100A7ox), Cys47 and Cys96 form an intramolecular disulfide bond, whereas these residues exist as free thiols in the reduced form (S100A7red). In this chapter, we provide a step-by-step protocol for the purification of S100A7ox and S100A7red that affords each protein in high yield and purity. In this procedure, S100A7 is expressed in Escherichia coli BL21(DE3), and the homodimer is obtained following ammonium sulfate precipitation, folding, and column chromatography. Treatment of S100A7 with 1,4-dithiothreitol (DTT) affords S100A7red. A Cu(II)-catalyzed oxidation reaction is employed to obtain S100A7ox. A RP-HPLC method that allows for baseline separation of S100A7ox and S100A7red is provided, as well as a biochemical Zn(II)-binding assay that can be employed to evaluate the functional integrity of S100A7.