Τίτλος:
Molecular characterization of recombinant Mus a 5 allergen from banana fruit
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
Allergy to banana fruit appears to have become an important cause of fruit allergy in Europe. Among five allergens that have been found, beta-1,3-glucanase denoted as Mus a 5 was identified as a candidate allergen for the component-resolved allergy diagnosis of banana allergy. Because of the variations in protein levels in banana fruit, in this study Mus a 5 was produced as a fusion protein with glutathione-S-transferase in Escherichia coli. The recombinant Mus a 5 was purified under native conditions by a combination of affinity, ion-exchange, and reversed phase chromatography. N-terminal sequence was confirmed by Edman degradation and 55 % of the primary structure was identified by mass fingerprint, while the secondary structure was assessed by circular dichroism spectroscopy. IgG reactivity of recombinant protein was shown in 2-D immunoblot with anti-Mus a 5 antibodies, while IgG and IgE binding to natural Mus a 5 was inhibited with the recombinant Mus a 5 in immunoblot inhibition test. IgE reactivity of recombinant Mus a 5 was shown in ELISA within a group of ten persons sensitized to banana fruit. Recombinant Mus a 5 is a novel reagent suitable for the component-resolved allergy diagnosis of banana allergy. © 2013 Springer Science+Business Media.
Συγγραφείς:
Mrkic, I.
Abughren, M.
Nikolic, J.
Andjelkovic, U.
Vassilopoulou, E.
Sinaniotis, A.
Petersen, A.
Papadopoulos, N.G.
Gavrovic-Jankulovic, M.
Περιοδικό:
Molecular Biotechnology
Εκδότης:
Humana Press Inc.
Λέξεις-κλειδιά:
Allergens; Circular dichroism spectroscopy; Escherichia coli; Fruits; Ion exchange; Proteins, Food allergies; Glucanase; Glutathione-S-transferase; IgE reactivities; Molecular characterization; Mus a 5; Reversed phase chromatography; Secondary structures, Allergies, allergen; immunoglobulin E; immunoglobulin G; recombinant allergen; recombinant mus a 5; recombinant protein; unclassified drug; allergen; glucan 1,3 beta glucosidase, affinity chromatography; amino acid sequence; amino terminal sequence; article; banana; circular dichroism; degradation kinetics; enzyme linked immunosorbent assay; immunoblotting; immunoglobulin structure; immunoreactivity; ion exchange chromatography; molecular dynamics; nucleotide sequence; protein secondary structure; reversed phase liquid chromatography; animal; Europe; food allergy; fruit; genetics; human; immunology; mouse; Musa, Escherichia coli, Allergens; Amino Acid Sequence; Animals; Europe; Food Hypersensitivity; Fruit; Glucan 1,3-beta-Glucosidase; Humans; Mice; Musa; Recombinant Proteins
DOI:
10.1007/s12033-013-9719-8
