Identification of the functional interleukin-2 binding domain of the chicken common cytokine receptor gamma chain.
CD132 is the common gamma chain to a number of cytokine receptor complexes including that for IL-2. To identify the functional domain of chicken CD132 (chCD132), the cDNA of chCD132 was cloned, and a neutralizing monoclonal antibody, raised against a recombinant chCD132 protein, was identified by inhibition of IL-2-dependent proliferation of T cells. Flow cytometry analysis revealed that chCD132 molecules are expressed on the surface of splenic mononuclear cells. The functional domain of chCD132 that binds to chicken interleukin 2, Q(84)E(94)L(95)Q(96)N(97)L(98), was found through phage display and peptide-competitive ELISA, and its critical residue Q(96) was further identified. A tertiary structure model shows that the functional domain is positioned at the elbow-like junction of N and C terminal fibronectin-III domains of chCD132. These data provide experimental evidence for elucidating the interaction between chCD132 and chIL-2.
Gu
Jianyou
J
Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou 310029, PR China.
Teng
Qiaoyang
Q
Huang
Zhenyu
Z
Ruan
Xizhen
X
Zhou
Jiyong
J
D000595:Amino Acid Sequence; D000818:Animals; D002469:Cell Separation; D002645:Chickens / Q000235:genetics* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005434:Flow Cytometry; D015870:Gene Expression; D020869:Gene Expression Profiling; D053631:Interleukin Receptor Common gamma Subunit / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007376:Interleukin-2 / Q000235:genetics* / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D053631:Interleukin Receptor Common gamma Subunit; D007376:Interleukin-2; D019151:Peptide Library; D011994:Recombinant Proteins