重组人骨唾液酸蛋白的纯化及生物学活性研究  

Study on purification and bioactivity analysis of recombinant human bone sialoprotein expressed by Pichia pastoris

作　　者：董燕[1] 王捷[1] 谭刘欣[1] 张宏斌[1] 武婕[1] 

机构地区：[1]广州军区广州总医院医学实验中心

出　　处：《中国生化药物杂志》2006年第2期71-74,78,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：广东省自然科学基金研究团队项目(No.20023001);广东省自然科学基金自由申请项目(No.04003534)

摘　　要：目的对毕赤酵母表达的重组人骨唾液酸蛋白(rhBSP)进行纯化和活性研究,为进一步研究BSP功能活性,揭示相关病理机制奠定基础。方法毕赤酵母(Pichia pastoris)GS115/pPICZαA-hbsp工程菌能高效分泌表达rhB-SP,采用Ni-NTA介质以亲和色谱法纯化rhBSP。通过羟基磷灰石(HA)晶体生长抑制实验、细胞黏附实验和鸡胚尿囊绒膜实验分析rhBSP的生物学活性。结果纯化的rhBSP达到电泳纯,并能抑制HA晶体生长,可介导乳腺癌细胞黏附和促进血管生成。结论纯化的rhBSP具有良好的生物学活性,可用于进一步研究BSP相关的病理、生理机制。Purpose To purify recombinant human bone sialoprotein （rhBSP） expressed by Pichia pastoris and to analyse its bioactivity so as to provide a basis for the deep research on biology and pathology of BSP. Methods Recombinant Pichia pastoris GSll5/pPICZaA-hbsp can express and secrete rhBSP highly, rhBSP was purified by immobilized metal ion affinity chromatography （IMAC） through Ni-NTA column. The bioactivity of hydroxyapatite seed growth inhibition, cell adhesion and angiogenesis was analyzed. Results rhBSP purified can inhibit the growth of hydroxyapatite seed, mediate breast cancer cells attachment and promote angiogenesis. Conclusion rhBSP has good bioactivity, and can be used to deeply study the pathology and physiology function of BSP.

关 键 词：人骨唾液酸蛋白 毕赤酵母 表达 纯化 活性 

分 类 号：R730.2[医药卫生—肿瘤] R392.11[医药卫生—临床医学]