The major human structural IgE epitope of the Brazil nut allergen Ber e 1: a chimaeric and protein microarray approach.
A protein microarray system containing different dilutions of 77 related and non-related proteins was used to show that IgE from subjects allergic to Brazil nut specifically recognise the seed 2S albumin protein (Ber e 1). Further, correctly folded chimaeric 2S albumin proteins containing structural epitope replacement were constructed and directed to the secretion pathway of the methylotropic yeast Pichia pastoris. Through the use of a chimaeric protein microarray system together with sera from a panel of 18 well-characterised Brazil nut allergic subjects, a structural IgE epitope of Ber e 1 was mapped to a helix-loop-helix region. The same structural region has been previously reported as the immunodominant region in related food allergens by different techniques. In conclusion, the combination of chimaeric proteins and protein microarrays will greatly facilitate the screening of a large number of individuals for a particular structural epitope and help to further our understanding of how proteins are recognised by the adaptive immune system.
School of Biosciences, Division of Nutritional Biochemistry, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, UK. marcos.alcocer@nottingham.ac.uk
Marcos J C
MJ
Alcocer
Gareth J
GJ
Murtagh
Philip B
PB
Wilson
Pavlos
P
Progias
Jing
J
Lin
David B
DB
Archer
D055516:2S Albumins, Plant; D000418:Albumins / Q000737:chemistry* / Q000276:immunology*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D031383:Bertholletia / Q000276:immunology*; D002942:Circular Dichroism; D000939:Epitopes; D005512:Food Hypersensitivity; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000276:immunology*; D040081:Protein Array Analysis; D017510:Protein Folding; D011498:Protein Precursors / Q000737:chemistry* / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D016415:Sequence Alignment
D055516:2S Albumins, Plant; D000418:Albumins; D052179:Antigens, Plant; D000939:Epitopes; D010455:Peptides; D010940:Plant Proteins; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C103364:2S albumin, brazil nut; D007073:Immunoglobulin E