可结合细胞表面IL-2Rα的短肽分子的筛选  

Screening of short peptides binding to cell surface interleukin-2 receptor α chain

作　　者：刘北一[1] 朱平[1] 罗海波[1] 富宁[1] 

机构地区：[1]南方医科大学免疫学教研室,广东广州510515

出　　处：《南方医科大学学报》2006年第7期971-974,共4页Journal of Southern Medical University

基　　金：广东省自然科学基金(32877)~~

摘　　要：目的利用噬菌体肽库技术筛选可特异性结合IL-2Rα的短肽序列,探索获得IL-2Rα小分子拮抗剂的可能性。方法以高表达IL-2Rα的MT-2细胞为钓饵,筛选噬菌体线性12肽库,用抗IL-2Rα单克隆抗体竞争洗脱结合的噬菌体,细胞ELISA、免疫组化鉴定阳性噬菌体克隆,并测序。结果随机挑选的17个克隆中有7个可与MT-2细胞结合。免疫组化显示阳性噬菌体克隆M15可与MT-2细胞及PHA刺激的PBMC结合。根据阳性克隆DNA序列,推导出氨基酸序列,共有6种序列,富含亲水氨基酸并包含Tyr、Phe、Leu保守残基。结论得到含Tyr、Phe保守残基的序列,阳性序列可结合细胞表面的IL-2Rα。Objective To screen and characterize the short peptides which bind specifically to interleukin-2 （IL-2） receptor a chain （IL-2Rα） for acquisition of small antagonists for blocking the binding of IL-2 with IL-2Rα. Methods 12-mer phage displayed peptide library was screened with the target cells of MT-2 cells which expressed IL-2Rα at high levels. The binding phage clones were eluted by anti-IL-2Rα monoclonal antibody. After 3 rounds of screening, the positive phage clones were identified by enzyme-linked immunosorbent assay （ELISA） and immunohistochemistry, and the amino acid sequences of the positive clones were deduced from the DNA sequences. Results Seven positive clones were screened out of the 17 phage clones bound to MT-2 cells. The positive clone M15 could bind specifically to MT-2 cell and PHA-activated peripheral blood monouclear cells. Amino acid sequence analysis identified 6 sequences, all of which contained hydrophilic residues, and 5 of these 6 sequences included Tyr, Phe and Leu conservative residues. Conclusion The peptide sequences containing Tyr, Phe conservative residues identified in this studv can bind to cell surface IL-2Rα.

关 键 词：噬菌体肽库 短肽序列 IL-2RΑ IL-2 

分 类 号：R392[医药卫生—免疫学]