Recombinant allergen Lol p II: expression, purification and characterization
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- 作者：Tamborini ; Elena ; Brandazza ; Anna ; de Lalla ; Claudia ; Siccardi ; Antonio G. ; Sidoli ; Alessandro ; et. al.
- 关键词：allergens ; perennial rye grass ; IgE ; recombinant proteins
- 刊名：Molecular Immunology
- 出版年：1995
- 出版时间：May, 1995
- 年：1995
- 卷：32
- 期：7
- 页码：505-513
- 全文大小：1.24 M
文摘
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 recombinantLol 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 naturalLol 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 recombinantLol p II is a highly stable and soluble monomeric molecule which behaves like a small globular protein.