摘要
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 (P2 2肽 ) ,经高效液相色谱纯化 ,毛细管电泳鉴定。结果 :合成多肽 (P2 2 )的纯度为 95 19% ,每次缩合的平均效率为 99 78% ;能与anti C5aRMcAb(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C2 0 ;还可被配体rh C5a( 10ng ml)明显抑制而降低其酶联OD值 (P <0 0 5 ) ,此外 10ng mlP2 2还可抑制rhC5a所致U937细胞胞浆Ca2 + 升高 (P <0 0 1)。结论 :从C5aR分子中寻找C5a结合位基序 ,可拮抗C5a的过敏毒素作用 。
Objective:To discover some high hydrophilic profiles of the human C5a anaphylatoxin based on relationship between the structure and function of the protein and the protein molecular design principles.Methods:The peptides were synthesized by 431A automatic peptide synthesizer,purified by PHLC and 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 Ca 2+ concentration was inhibited by P22 in dt2cAMP differentiated 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.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2001年第3期125-129,共5页
Chinese Journal of Immunology
基金
自然科学基金资助! (编号 :39770 315
39970 330 )