title: Pro j 2 is mesquite profilin: molecular characteristics and specific IgE binding activity.
abstract: BACKGROUND
  Pollens from mesquite (Prosopis juliflora) are potent allergen responsible in causing immediate hypersensitivity reactions in susceptible people in tropical countries.
  
  
  OBJECTIVE
  This study aimed to clone, express and purify the mesquite pollen profilin (Pro j 2) as well as evaluating its nucleotide sequence homology in order to predict allergenic cross-reactivity with profilins of common allergenic plants.
  
  
  METHODS
  Immunoblotting assay and specific ELISA were applied to determine the immunoreactivity of sera from 35 patients who were allergic to mesquite pollen. The mesquite profilin-coding sequence was cloned into PTZ57R/T vector and amplified. The cDNA of mesquite pollen profilin was then expressed in Escherichia coli using pET-21b (+) vector and puri?ed by one-step Ni2+ a?nity chromatography. IgE binding capacity of the recombinant mesquite profiling (rPro j 2) was analyzed by specific ELISA, immunoblotting, and inhibition assays.
  
  
  RESULTS
  cDNA nucleotide sequencing revealed an open reading frame of 399bp encoding for 133 amino acids which belongs to the profilin family. Seventeen patients (17/35, 48.57%) had significant specific IgE level for rPro j 2. Immunodetection and inhibition assays indicated that puri?ed rPro j 2 might be similar as that in the crude extract.
  
  
  CONCLUSIONS
  Pro j 2, as a new allergen from mesquite pollen, was produced in E. coli with an IgE-reactivity similar to that of its natural counterpart. The amino acid sequences homology analysis of mesquite profilin and several profilin molecules from other plants showed high degree of cross-reactivity among plant-derived profilins from unrelated families.
authors.affiliation: Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: Hosein
authors.initials: H
authors.lastname: Ali-Sadeghi
authors.forename: Ali
authors.initials: A
authors.lastname: Khodadadi
authors.forename: Akram
authors.initials: A
authors.lastname: Amini
authors.forename: Mohammad-Ali
authors.initials: MA
authors.lastname: Assarehzadegan
authors.forename: Najmeh
authors.initials: N
authors.lastname: Sepahi
authors.forename: Farnoosh
authors.initials: F
authors.lastname: Zarinhadideh
mesh_terms: D000595:Amino Acid Sequence; D052179:Antigens, Plant / Q000009:adverse effects / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001667:Binding, Competitive; D016022:Case-Control Studies; D003001:Cloning, Molecular; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology*; D008297:Male; D010940:Plant Proteins / Q000009:adverse effects / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011058:Pollen / Q000009:adverse effects / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051304:Profilins / Q000009:adverse effects* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D029862:Prosopis / Q000009:adverse effects / Q000235:genetics / Q000276:immunology*; D011485:Protein Binding; D011994:Recombinant Proteins; D006255:Rhinitis, Allergic, Seasonal / Q000097:blood / Q000276:immunology* / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017385:Sequence Homology
chemical_list: D052179:Antigens, Plant; D010940:Plant Proteins; C000613609:Pro j 2 allergen, Prosopis juliflora; D051304:Profilins; D011994:Recombinant Proteins; D007073:Immunoglobulin E