蓝贻贝抗凝肽的合成及活性初步研究

Preliminary study on synthesis and activity of blue mussel anticoagulant peptide

目的采用多肽固相合成法合成蓝贻贝抗凝肽MEAP(Blue mussel Mytilus edulis anticoagulant peptide),初步研究其抗凝活性。方法以2-CTC树脂为载体,Fmoc保护氨基酸为原料,按照蓝贻贝抗凝肽的序列从C端向N端逐步偶联氨基酸,裂解后得到粗肽,经半制备液相纯化后测定精肽的凝血酶时间(TT)、活化的部分凝血活酶时间(APTT)、凝血酶原时间(PT)。结果合成的蓝贻贝抗凝肽粗品的纯度为65.5%,经半制备纯化后的精肽纯度为95.5%;抗凝活性实验显示APTT、TT具有显著的延时作用(P<0.01)。结论固相合成的蓝贻贝抗凝肽具有抗凝活性。

Objective To prepare blue mussel Mytilus edulis anticoagulant peptide （MEAP） by using sol- id phase peptide synthesis method and to preliminarily study its anticoagulant activity. Methods MEAP was successfully synthesized from its C terminal to N terminal according to its peptide sequence by u- sing 2-CTC resin as the solid supporter and Fmoc amino acids as raw materials. The crude peptide was cleaved from the resin and purified by RP-HPLC. The thrombin time （TT）, activation of blood coagu- lation time （APTT） and prothrombin time （PT） of purified MEAP were determined. Results The puri- ties of synthetic crude and purified MEAP were 65.5 % and 95.5%, respectively. Anticoagulant activi- ty experiments showed that both APTT and TT of synthetic MEAP had significant delayed effect （P〈0.01）. Conclusion MEAP obtained by using solid phase peptide synthesis method had anticoagulant ac- tivity.