中国寄生虫学与寄生虫病杂志 ›› 1999, Vol. 17 ›› Issue (4): 12-230.

• 论著 • 上一篇    下一篇

伯氏疟原虫氯喹结合蛋白的分离与定量

王琴美1,王鸣杰1,常惠玲 1,杨彬2

  

  1. 中国预防医学科学院寄生虫病研究所 上海 200025;南京药物研究所 南京 210009
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1999-08-30 发布日期:1999-08-30

ISOLATION AND QUANTITATION OF CHLOROQUINE-BINDING PROTEINS IN PLASMODIUM BERGHEI *

WANG Qinmei 1;WANG Mingjie 1;CHANG Huiling 1;YANG Bin 2   

  1. 1 Institute of Parasitic Diseases **;Chinese Academy of Preventive Medicine Shanghai;Shanghai 200025 2 Nanjing Institute of Materia Medica;Nanjing 210009
  • Received:1900-01-01 Revised:1900-01-01 Online:1999-08-30 Published:1999-08-30

摘要:   目的: 分离伯氏疟原虫敏感株( C S) 与抗氯喹株的氯喹(chloroquine C Q) 结合蛋白。方法: 给两株感染鼠ig C Q400 mg/kg , 3 h 后, 收集伯氏疟原虫用( Ultrogel R) Ac A34 凝胶柱分离蛋白, 按 Bergqvist 法抽提与蛋白结合的 C Q, 然后用 H P L C ( 外标法) 定量。结果: C S 有58 个蛋白峰, 均与 C Q 结合, 其中以 No64 ~86部分( 峰9) 结合的 C Q 量最多, 占总结合量的164 % 。抗氯喹伯氏疟原虫( C R) 有40 个蛋白峰, 除8 个峰外,均与 C Q 结合, 结合量最多的蛋白部分与 C S相同, 在 No60 ~83 部分( 峰6 及峰7) , 但是它与 C Q 的结合能力( C Qnmol/ mg 蛋白) 显著高于 C S。结论: 分离与测定了 C S和 C R 两株疟原虫各个蛋白峰的分布与定量, 及其与 C Q 结合的能力。在同一株疟原虫中, 各蛋白与 C Q 的结合能力存有差异, 在 C S与 C R 间此差异更大。

关键词: 伯氏疟原虫, 蛋白峰, 蛋白结合能力, AcA_(34)柱, HPLC

Abstract:  AIM: To isolate and quantitate chloroquine (CQ)-binding proteins in both CQ-sensitive (CS) AND CQ-resistant (CR) Plasmodium berghei . METHODS: P.berghei infected mice were each given ig CQ 400 mg/kg, respectively, 3 h later, the proteins of the parasites collected were separated using Ultrogel R AcA 34 gel column , followed by CQ extraction , the CQ concentration was determined by HPLC (extra standard). RESULTS: There were 58 protein peaks in the CS, all of which were bound with CQ,fraction Nos. 64~86 (peak 9) being the predominant, accounting for 16.4% of the total amount of CQ-binding protein. In the CR there were 40 protein peaks, among which 32 were bound with CQ, the predominant fraction Nos.60~83 (peaks 6,7),had a higher CQ-binding ability than CS. CONCLUSION: The distribution and amount of each CQ-binding protein peak in both CS and CR strains of P.berghei were demonstrated, exhibiting differences in the ability of CQ-binding not only between CS and CR strains but also within the same strain.

Key words: Plasmodium berghei, protein peak, chloroquine binding ability, AcA 34 column, HPLC