Title
The allergen Mus m 1.0102: Dissecting the relationship between molecular conformation and allergenic potency
Author
Elena Ferrari, Emanuela Casali, Samuele E. Burastero, Alberto Spisni, Thelma A. Pertinhez
Year
2016
Journal
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics
Abstract
The species Mus musculus experiences an obligate proteinuria: predominant are the Major Urinary Proteins (MUPs), that, collectively known as the major mouse allergen Mus m 1, are among the most important aeroallergens for mouse allergic patients. The production of a soluble and stable hypoallergenic form of Mus m 1 is essential for the development of immunotherapeutic protocols to treat allergic symptoms. We introduced the substitution C138S in recombinant Mus m 1.0102, an allergenic isoform of Mus m 1. Solubility, conformation, stability and ability to refold after chemical denaturation were investigated with dynamic light scattering, circular dichroism, fluorescence and NMR spectroscopy. An in vitro degranulation assay was used to evaluate the protein allergenic potential, and compare it with Mus m 1.0102 and with an hypoallergenic variant bearing the substitution Y120A. Mus m 1.0102-C138S retains a native-like fold revealing, however, local conformational alterations that influence some of its physical and allergenic properties: it is monodispersed, thermostable up to 56 °C, able to reversibly unfold and it exhibits an enhanced allergenicity. The unique free thiol group affects the solution structural stability of the native protein. Because the mutant C138S does not aggregate over time it is a good lead protein to develop diagnostic and therapeutic applications.
Instrument
J-720
Keywords
Circular dichroism, Secondary structure, Tertiary structure, Protein denaturation, Protein folding, Biochemistry