Characterization and crystallization of a recombinant IgE Fab fragment in complex with the bovine beta-lactoglobulin allergen.
A D1 Fab fragment containing the allergen-binding variable domains of the IgE antibody was characterized by ESI FT-ICR mass spectrometry and crystallized with bovine beta-lactoglobulin (BLG) using the hanging-drop vapour-diffusion method at 293 K. X-ray data suitable for structure determination were collected to 2.8 A resolution using synchrotron radiation. The crystal belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 67.0, b = 100.6, c = 168.1 A. The three-dimensional structure of the D1 Fab fragment-BLG complex will provide the first insight into IgE antibody-allergen interactions at the molecular level.
Niemi
Merja
M
Department of Chemistry, University of Joensuu, PO Box 111, FIN-80101 Joensuu, Finland. merja.niemi@joensuu.fi
Jänis
Janne
J
Jylhä
Sirpa
S
Kallio
Johanna M
JM
Hakulinen
Nina
N
Laukkanen
Marja-Leena
ML
Takkinen
Kristiina
K
Rouvinen
Juha
J
D000485:Allergens / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002648:Child; D018360:Crystallography, X-Ray; D005512:Food Hypersensitivity / Q000276:immunology; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000737:chemistry*; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry; D007147:Immunoglobulin Light Chains / Q000737:chemistry; D007782:Lactoglobulins / Q000737:chemistry* / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology; D008969:Molecular Sequence Data; D013057:Spectrum Analysis
D000485:Allergens; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007782:Lactoglobulins; D007073:Immunoglobulin E