人血清载脂蛋白H的纯化与鉴定  

Purification and Identification of Human Serum Apolipoprotein H

作　　者：蒋磊[1] 郑颂阳[1] 石庆杰[1] 陈检芳[1] 王克超[1] 刘健平[1] 程冠生[1] 

机构地区：[1]湖南医科大学同位素室,湖南长沙410078

出　　处：《标记免疫分析与临床》2003年第1期29-32,共4页Labeled Immunoassays and Clinical Medicine

摘　　要：为了研究载脂蛋白H(apoH)与疾病的关系 ,分别用HClO4,(NH4) 2 SO4沉淀、DEAE -纤维素离子交换层析 ,从人血清中纯化了载脂蛋白H。用测定分子量 ,分析氨基酸组成和N -端氨基酸测序三个方法进行鉴定。用SDS -PAGE测得其相对分子量为 5 4 0 0 0。用高效液相色谱测定其中 15种氨基酸的相对含量 (mol/ 10 3 mol残基 )如下 :Asp119.8,Glu10 8,Thr94 .8,Ser78.6 ,Phe6 4 .9,Gly10 0 8,Ala5 7.2 ,Val5 7.2 ,Met10 .6 ,Ile5 0 .3,Leu77.1,Tyr31.9,His15 .2 ,L - ys81.4 ,Arg39.4 ;在PE -ABI公司生产的 4 75A型气相蛋白质测序仪上测得其N端 10个氨基酸残基顺序如下 :NH2 -Gly -Arg -Thr -Cys -Pro -Lys -Pro -Asp -Asp -Leu。纯化得到的apoH与肝素有高的亲和性 ,还能与乙肝表面抗原结合。纯化apoH的方法相对比较简单 。In order to study the relationships between apoH and diseases, apoH was purified from human serum by fractionation with HClO 4, (NH 4) 2SO 4 followed by chromatography on DEAE-cellulose-32 ion exchange column. Considering the characteristics of human serum apoH and various conditions in our laboratory, the purified protein was identified by determining its molecular weight and amino acid components and sequencing its partial amino acid residues at the N-terminal region. The protein was estimated to be 54000 by SDS-PAGE; the percentages of 15 amino acids were as follows (mol/10 3mol residue) : Asp119.8, Glu108, Thr94.8, Ser78.6, Phe64.9, Gly1008, Ala57,2, Val57.2, Met10.6, Ile50.3, Leu77.1, Tyr 31.9, His15.2, Lys81.4, Arg39,4; the sequence of its 10 amino acids at the N-terminal region measured on 475A model protein sequencer was as follows: NH 2-Gly-Arg-Thr-Cys-Pro-Lys-Pro-Asp-Asp-Leu. In addition, this protein had an affinity with heparin, and it also has an ability to bind to hepatitis B virus surface antigen. All above results are consistent with those of human serum apoH reported in literatures. The methods used here to purify human serum apoH are relatively simple, and the purified apoH can be used to the protein sequencing. success of purification of human apoH lays the foundation for preparation of apoH determining test kit.

关 键 词：人血清 载脂蛋白H 纯化 鉴定 

分 类 号：Q503[生物学—生物化学]