Effect of Point Mutations on Structural and Allergenic Properties of the Lentil Allergen Len c 3

Membranes

Natalia S. Matveevskaya¹  Ivan V. Bogdanov²  Tatiana V. Ovchinnikova²  Anastasia A. Ignatova²  Andrey A. Tagaev²  Daria N. Melnikova²  Ekaterina I. Finkina² 

[1] G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology, Admiral Makarov St., 10, 125212 Moscow, Russia;M.M. Shemyakin & Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, The Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, 117997 Moscow, Russia;

关键词: Len c 3;    site-directed mutagenesis;    recombinant analogue;    allergen;    IgE-binding capacity;    lipid binding;   

DOI  :  10.3390/membranes11120939

来源: DOAJ

【 摘 要 】

Plant lipid transfer proteins (LTPs) are known to be clinically significant allergens capable of binding various lipid ligands. Recent data showed that lipid ligands affected the allergenic properties of plant LTPs. In this work, we checked the assumption that specific amino acid residues in the Len c 3 structure can play a key role both in the interaction with lipid ligands and IgE-binding capacity of the allergen. The recombinant analogues of Len c 3 with the single or double substitutions of Thr41, Arg45 and/or Tyr80 were obtained by site-directed mutagenesis. All these amino acid residues are located near the “bottom” entrance to the hydrophobic cavity of Len c 3 and are likely included in the IgE-binding epitope of the allergen. Using a bioinformatic approach, circular dichroism and fluorescence spectroscopies, ELISA, and experiments mimicking the allergen Len c 3 gastroduodenal digestion we showed that the substitution of all the three amino acid residues significantly affected structural organization of this region and led both to a change of the ligand-binding capacity and the allergenic potential of Len c 3.

【 授权许可】

Unknown