Recombinant allergen Lol p II: expression, purification and characterization.
Pollen from perennial rye grass (Lolium perenne) is a major cause of type I allergies worldwide. It contains complex mixtures of proteins, among which Lol p II is a major allergen. Previously, we have reported the cloning and sequencing of Lol p II and its expression in fusion with the heavy chain of human ferritin as carrier polypeptide (Sidoli et al., 1993, J. biol. Chem. 268, 21819-21825). Here, we describe the expression, purification and characterization of a recombinant Lol p II overproduced as a non-fusion protein in the periplasm of E. coli. The recombinant allergen was expressed in high yields and was easily purified in milligram amounts. It competed with the natural Lol p II for binding to specific IgE, and it induced allergic responses in skin prick tests, indicating to be immunologically analogous to the natural protein. Biochemical analyses indicate that recombinant Lol p II is a highly stable and soluble monomeric molecule which behaves like a small globular protein.
Tamborini
E
E
Department of Biology and Technology, San Raffaele Scientific Institute, Milano, Italy.
Brandazza
A
A
De Lalla
C
C
Musco
G
G
Siccardi
A G
AG
Arosio
P
P
Sidoli
A
A
D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008129:Lolium / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011058:Pollen / Q000737:chemistry / Q000235:genetics*; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D006255:Rhinitis, Allergic, Seasonal / Q000175:diagnosis / Q000276:immunology; D012882:Skin Tests
D000485:Allergens; D052179:Antigens, Plant; D017931:DNA Primers; C056754:Lol p II protein, Lolium perenne; D010940:Plant Proteins; D011994:Recombinant Proteins