Identification of functional domains in goose interleukin 2.
The cytokine interleukin (IL)-2 functions as a growth factor and central regulator in the immune system. Using a recombinant goose IL-2 (goIL-2) monomer expressed in prokaryotic cells as an immunogen, we synthesized 5 goIL-2 neutralizing mAbs to identify the functional domains of goIL-2, and used these mAbs to finely map the functional domains of the goIL-2 protein. The mimotopes of the 5 anti-goIL-2 mAbs, including HHDPWDXLP, ESLSRXXMXXLXP, SHHLPTSXL, HPDPWDAPLSS, and HEPWQLXL, were identified using a phage display library and peptide-competitive enzyme-linked immunosorbent assay (ELISA). These mimotopes constitute 1 conformational functional domain in the goIL-2 molecule--T¹¹I¹⁴K¹⁵D¹⁶E¹⁸K¹⁹L²⁰G²¹T²²S²³M²⁴K²⁵L²⁹E³⁰L³¹Y³²T³³P³⁴E³⁶S⁴¹W⁴²Q⁴³T⁴⁴L⁴⁵Q⁴⁶ (domain I). The neutralizing mAbs to goIL-2 inhibited the in vitro lymphocyte proliferation stimulated by domain I peptides of goIL-2. A tertiary structural model of goIL-2 showed that domain I is positioned in helix A, long A-B loop, and the N-terminal of helix B. These data provide a clue for defining the interaction between goIL-2 and its receptor.
Ma
Guangpeng
G
Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou 310029, PR China.
Chen
Xinyu
X
Gu
Jianyou
J
Zhou
Jiyong
J
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D030161:Avian Proteins / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D005777:Geese / Q000235:genetics* / Q000276:immunology*; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000737:chemistry* / Q000235:genetics*; D008213:Lymphocyte Activation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D030161:Avian Proteins; D000939:Epitopes; D007376:Interleukin-2; D019151:Peptide Library; D011994:Recombinant Proteins