化学学报 ›› 2004, Vol. 62 ›› Issue (3): 279-282. 上一篇    下一篇

研究论文

Ca2+与乳清蛋白结合的亲和毛细管电泳研究

王畅, 郑育芳, 熊建辉, 翁前锋, 杨军, 杨青, 许国旺   

  1. 中国科学院大连化学物理研究所国家色谱分析中心, 大连, 116011
  • 投稿日期:2003-07-04 修回日期:2003-10-08 发布日期:2014-01-26
  • 通讯作者: 许国旺,E-mail:dicp402@mail.dlptt.ln.cn;Tel:(0411)3693403. E-mail:dicp402@mail.dlptt.ln.c
  • 基金资助:
    中国科学院领域前沿创新基金(Nos.K2002A11,K2002A12)资助项目.

Study on Human α-Lactalbumin-Calcium Interaction Using Affinity Capillary Electrophoresis

WANG Chang, ZHENG Yu-Fang, XIONG Jian-Hui, WENG Qian-Feng, YANG Jun, YANG Qing, XU Guo-Wang   

  1. National Chromatographic Research & Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011
  • Received:2003-07-04 Revised:2003-10-08 Published:2014-01-26

利用亲和毛细管电泳研究了Ca2+与α-人乳清蛋白(α-HLA)的结合情况.以恒定浓度α-HLA作为受体,运行缓冲溶液加入不同浓度的Ca2+作为配体,可观察到由于Ca2+的结合,α-HLA的电泳淌度发生了变化.通过Scatchard方程的淌度比(M)处理数据得到α-HLA与Ca2+的表观结合常数(Kapp)为2.0×107(mol/L)-1.同时考察了Ca2+对变性剂(尿素)和热诱导所引起的α-HLA去折叠的影响,结果表明,Ca2+的结合增强了α-HLA的稳定性,也即提高了α-HLA抗变性剂和热诱导的去折叠性能.

关键词: 亲和毛细管电泳, α-人乳清蛋白, Ca2+结合

Affinity capillary electrophoresis was used to study the interaction of human α-lactalbumin(α-HLA)with calcium ion(Ca2+).With constant concentration ofα2HLA as a receptor and various concentrations of Ca2+ as a ligand in the running buffer,changes in electrophoretic mobilities ofα-HLA were observed when complexes of α-HLA and Ca2+ came into being.Based on Scatchard analysis of the mobility ratios(M)of α-HLA to Ca2+,it was found that the apparent binding constant(Kapp)was 2.0×107(mol/L)-1.In addition,the effect of Ca2+ on the unfolding ofα2HLA induced by urea and heating was studied,and the results showed that the binding of Ca2+ increased the stability of α-HLA against the action of denaturing agent(such as urea)and heating.

Key words: affinity capillary electrophoresis, human α-lactalbumin, calcium binding