露峰房抗炎蛋白中多肽成分的分离、纯化及性质研究

 目的:从药用露峰房(Nidus vespae)筛选出的具有较好的抗炎、免疫活性蛋白粗品——NV₃中分离纯化,得到一多肽类化合物,命名为NV-PP-1,并检测其理化性质?方法:以离子交换层析、排阻层析、HPLC等进行分离纯化,并以高效凝胶排阻色谱(HPSEC)、高效毛细管等电聚焦电泳(IEF-HPCE)、亲水性分子排阻液相Protein-Pak^TM60柱、氨基酸分析、DNS-Cl法的N-末端测定检测其理化性质。结果:高效凝胶排阻色谱(HPSEC)及高效毛细管等电聚焦电泳(IEF-HPCE)检测其皆为单峰。亲水性分子排阻液相Protein-Pak^TM60柱测得其分子量为7.079kD。IEF-HPCE确定其PI值为4.08。紫外吸收最高峰为274nm。氨基酸分析表明其含有近56个氨基酸残基,其中谷氨酸、丝氨酸、甘氨酸较多,还有天冬氨酸等,是一酸性多肽。末端测定表明其N-末端封闭。结论:酸性多肽NV-PP-1是从药用露蜂房抗炎蛋白粗品中首次分离得到的成分。

Isolation purification and characterization of NV-PP-1 from Nidus vespae

OBJECTIVES:To isolate and purify a new peptide named as NV PP 1,from anti inflammatory protein extracts of Nidus Vespae and to characterize it.METHODS:It was purified by ammonium sulfate precipitation,DEAE Sephadex A50 ionexchange chromatography,Sephadex G-50 gel filtration and HPLC,respectively.Then it was characterized by HPLC on Protein Pak TM 60 column,IEF HPCE(isoelectric focusing high performance capillary electrophoresis),UV spectrum,amino acid analysis,and the N terminal analysis with DNS Cl.RESULT:Both HPLC on Protein Pak TM 60 column and IEF HPCE showed its homogeneity.Its molecular weight was estimated to be 7.079 kD by HPSEC.UV spectrum indicated its maximum absorption at 274nm.Its PI was determined to be 4.08 according to IEF HPCE.The terminal analysis showed its N terminal amino acid was blocked.Amino acid analysis of NV PP 1 indicated that it was rich in Ser,Glu,Gly.CONCLUSION:NV PP 1 is the first peptide isolated from the anti inflammatory protein extracts of Nidus vespae.