人C5a过敏毒素拮抗剂的分子设计及其活性检测

目的：从蛋白质结构与功能的关系出发，探讨C5aR与其配体的C5a的结合位。方法：按分子设计理论，采用亲水性方案寻找C5aR胞外区高亲水性区域，Fmoc方案人工合成C5aR第9－30位氨基酸基序（P22肽），经高效液相色谱纯化，毛细管电泳鉴定。结果：合成多肽（P22）的纯度为95.19％，每次缩合的平均效率为99.78％；能与anti-C5aR McAb（S5／1，Serotic公司）有效地结合，酶联OD490极显著高于同浓度对照多肽C20；还可被配体rh-C5a(10ng/ml)明显抑制而降低其酶联OD值（P＜0.05），此外10ng/mlP22还可抑制rhC5a所致U937细胞胞浆Ca^2 升高（P＜0.01）。结论：从C5a分子中寻找C5a结合位基序，可拮抗C5a的过敏毒素作用，为治疗C5a及其相关疾病进行新型的药物设计。

Molecular design forantagonisting C5a anaphyl atoxin and the activity examination of the peptide

Objective:To discover some high hydrophilic profiles of the humanC5a anaphylatoxin based on relationship between the structure and function of the protein and the prot ein molecular design principles.Methods:The peptides were synthesized by 431A automatic pe ptide synthesizer,purified by PHLC an d confirmed by caplilliary electrophoresis.Results:The N- terminus No.9-30 profile of the C5aR(P22) could interacte with anti-C5aR McAb( S5/1,from Serotic Co.),as determined by ELISA.Furthermore,it could be inhibited OD490 values remarkably by 10.0 μg/L rhC5a(P＜0.05)as well.In addition,the cytoplasmic Ca2+ concentration was inhibited by P22 in dt2cAMP differentia ted U937 that induced by 10.0 μg/L rhC5a(P＜0.01).Conclusion:It is possible that the C5a anaphylatoxin would be removed from the body,and some newtype pharmaceuticals that therapy disease interrelated C5a anaphylatoxin could be manufactured.