Purification and characterisation of relevant natural and recombinant apple allergens.
Apple (Malus domestica) is the most widely cultivated fruit crop in Europe and frequently causes allergic reactions with a variable degree of severity. So far, four apple allergens Mal d 1, Mal d 2, Mal d 3 and Mal d 4 have been identified. Mal d 1, a Bet v 1 related allergen, and Mal d 4, apple profilin, are sensitive to proteolytic degradation, whereas Mal d 2, a thaumatin-like protein and Mal d 3, a nonspecific lipid transfer protein, are rather stable to proteolytic processes. Mal d 1 and Mal d 4 were purified after expression in Escherichia coli expression system, while Mal d 2 and Mal d 3 were purified from apple fruit tissue. All purified proteins were subjected to detailed physicochemical characterisation to confirm their structural integrity and maintained IgE binding capacity. Detailed investigations of carbohydrate moieties of Mal d 2 demonstrated their involvement in the overall IgE binding capacity of this allergen. It was concluded that the folded structure and IgE binding capacity of all four allergens were preserved during purification.
Oberhuber
Christina
C
Biomay, Vienna, Austria.
Ma
Yan
Y
Marsh
Justin
J
Rigby
Neil
N
Smole
Ursula
U
Radauer
Christian
C
Alessandri
Stefano
S
Briza
Peter
P
Zuidmeer
Laurian
L
Maderegger
Bernhard
B
Himly
Martin
M
Sancho
Ana I
AI
van Ree
Ronald
R
Knulst
André
A
Ebner
Christof
C
Shewry
Peter
P
Mills
E N Clare
EN
Wellner
Klaus
K
Breiteneder
Heimo
H
Hoffmann-Sommergruber
Karin
K
Bublin
Merima
M
D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D052179:Antigens, Plant / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002352:Carrier Proteins; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D027845:Malus / Q000276:immunology*; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
D000485:Allergens; D052179:Antigens, Plant; D002352:Carrier Proteins; C095551:MALD1 protein, Malus domestica; C531902:Mal d 2 allergen, Malus domestica; C463180:Mal d 3 allergen; C518790:Mal d 4 allergen, Malus domestica; D010940:Plant Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E