人血清载脂蛋白H的纯化与鉴定

为了研究载脂蛋白H（apoH)与疾病的关系，分别用HClO4,(NH4)2SO4沉淀、DEAE－纤维素离子交换层析，从人血清中纯化了载脂蛋白Ho用测定分子量，分析氨基酸组成和N－端氨基酸测序三个方面进行鉴定。用SDS－PAGE测得其相对分子量对54000。用高效液相色谱测定其中15种氨基酸的相对含量（mol/10^3mol残基）如下：Asp119.8,Glu108,Thr94.8,Ser78.6,Phe64.9,Gly1008,Ala57.2,Val57.2,Met10.6,Ile50.3,Leu77.1,Tyr31.9,His15.2,L-ys81.4,Arg39.4;d PE-ABI公司生产的475A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-Cys-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，能达到测序的要求。

Purification and Identification of Human Serum Apolipoprotein H

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.