Title
The major allergen Der p 2 is a cholesterol binding protein
Author
Kavita Reginald, Fook Tim Chew
Year
2019
Journal
Scientific Reports
Abstract
Der p 2 is a major dust mite allergen and >80% of mite allergic individuals have specifc IgE to this allergen. Although it is well characterized in terms of allergenicity, there is still some ambiguity in terms of its biological function. Three-dimensional structural analysis of Der p 2 and its close homologues indicate the presence of a hydrophobic cavity which can potentially bind to lipid molecules. In this study, we aimed to identify the potential ligand of Der p 2. Using a liposome pulldown assay, we show that recombinant Der p 2 binds to liposomes prepared with exogenous cholesterol in a dose dependent fashion. Next, an ELISA based assay using immobilized lipids was used to study binding specifcities of other lipid molecules. Cholesterol was the preferred ligand of Der p 2 among 11 diferent lipids tested. Two homologues of Der p 2, Der f 2 and Der f 22 also bound to cholesterol. Further, using liquid chromatography-mass spectrometry (LC-MS), we confrmed that cholesterol is the natural ligand of Der p 2. Three amino acid residues of Der p 2, V104, V106 and V110 are possible cholesterol binding sites, as alanine mutations of these residues showed a signifcant decrease in binding (p
Full Article
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Biochemistry