Identification of Cucumisin (Cuc m 1), a subtilisin-like endopeptidase, as the major allergen of melon fruit.
BACKGROUND
Allergenic components in melon extracts have not been described in spite of the fact that melon (Cucumis melo) is a frequent allergy-eliciting fruit. The aim of this study was to evaluate allergenic components in melon extract and to report the identification of cucumisin as a major melon allergen.


METHODS
Sera from 35 patients allergic to melon were selected on the basis of clinical symptoms, skin prick tests and oral challenge test. Allergenic components were detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting. Molecular characterization of IgE-binding bands was performed by N-terminal amino acid sequencing.


RESULTS
More than 10 IgE-binding bands, between 10 and 80 kDa, were identified in melon extract. Out of them, four IgE-binding bands were major allergens: 14 kDa, 36 kDa, 54 kDa and 67 kDa. These major allergens, except 14 kDa band, showed the same N-terminal sequence: T-T-R-S-W-D-F-L. Research conducted with protein databases identified this N-terminal sequence as cucumisin, an alkaline serine protease, which shares structural homology with microbial subtilisin. The molecular mass of the identified bands corresponds with different molecular forms of cucumisin produced during the processing or degradation of the enzyme: 67 kDa native cucumisin, 54 kDa mature cucumisin and 36 kDa NH2-terminal cucumisin fragment.


CONCLUSIONS
Cucumisin (Cuc m 1) and several N-terminal cucumisin fragments are the major allergens of melon. The ubiquitous distribution of this protein family (cucumisin-like proteases) in many plant species and its high structural similarity suggest its potential role as a new panallergen in plant foods.
Cuesta-Herranz
J
J
Servicio de Alergia, Fundación Jiménez Díaz and C.B.F. Leti, SA. Tres Cantos, Madrid, Spain. jcuesta@fjd.es
Pastor
C
C
Figueredo
E
E
Vidarte
L
L
De las Heras
M
M
Durán
C
C
Fernández-Caldas
E
E
de Miguel
J
J
Vivanco
F
F
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000032:analysis* / Q000235:genetics; D000595:Amino Acid Sequence; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D028462:Cucumis / Q000276:immunology*; D028464:Cucurbita / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005512:Food Hypersensitivity / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007073:Immunoglobulin E / Q000378:metabolism; D018551:Solanum lycopersicum / Q000276:immunology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010936:Plant Extracts / Q000276:immunology; D012697:Serine Endopeptidases / Q000032:analysis / Q000235:genetics; D012882:Skin Tests
D000485:Allergens; D010936:Plant Extracts; D007073:Immunoglobulin E; D012697:Serine Endopeptidases; C041541:cucumisin