Purification, crystallization and preliminary X-ray characterization of prunin-1, a major component of the almond (Prunus dulcis) allergen amandin.
The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond ( Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 A and belong to the tetragonal space group P4(1)22, with unit cell parameters of a = b = 150.912 A, c = 165.248 A. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.
Albillos
Silvia M
SM
Illinois Institute of Technology, National Center for Food Safety and Technology, 6502 South Archer Road, Summit-Argo, Illinois 60501, USA.
Jin
Tengchuan
T
Howard
Andrew
A
Zhang
Yuzhu
Y
Kothary
Mahendra H
MH
Fu
Tong-Jen
TJ
D000595:Amino Acid Sequence; D052179:Antigens, Plant / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D005512:Food Hypersensitivity; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D010695:Phlorhizin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D027861:Prunus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D016415:Sequence Alignment
D052179:Antigens, Plant; D010455:Peptides; C463779:amandin; C506622:prunin; D010695:Phlorhizin