AbstractMalasseziaare the dominant commensal yeast species of the human skin microbiota and are associated with inflammatory skin diseases, such as atopic eczema (AE). The Mala s 1 allergen ofMalassezia sympodialisis a β-propeller protein, inducing both IgE and T-cell reactivity in AE patients. We demonstrate by immuno-electron microscopy that Mala s 1 is mainly located in theM. sympodialisyeast cell wall. An anti-Mala s 1 antibody did not inhibitM. sympodialisgrowth suggesting Mala s 1 may not be an antifungal target.In silicoanalysis of the predicted Mala s 1 protein sequence identified a motif indicative of a KELCH protein, a subgroup of β-propeller proteins. To test the hypothesis that antibodies against Mala s 1 cross-react with human skin (KELCH) proteins we examined the binding of the anti-Mala s 1 antibody to human skin explants and visualized binding in the epidermal skin layer. Putative human targets recognized by the anti-Mala s 1 antibody were identified by immunoblotting and proteomics. We propose that Mala s 1 is a KELCH-like β-propeller protein with similarity to human skin proteins. Mala s 1 recognition may trigger cross-reactive responses that contribute to skin diseases associated withM. sympodialis.