姜黄素衍生物C085与Hsp90及其不同片段相互作用以及对Hsp90 ATPase活性的影响  

Interactions between curcumin derivatives C085 and various constructs of Hsp90 and effects of C085 on Hsp90 ATPase activity

作　　者：范莹娟[1,2] 张连茹[3] 刘洋[1,2] 许建华[1,2] 

机构地区：[1]福建医科大学药学院 [2]福建省天然药物药理学重点实验室,福建福州350004 [3]厦门大学生命科学学院,福建厦门361005

出　　处：《中国药理学通报》2014年第10期1408-1413,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81173096);科技部"重大新药创制"科技重大专项(No 2012ZX09103-101-028)

摘　　要：目的：研究姜黄素衍生物C085与Hsp90的结合作用,及其对Hsp90-ATPase 活性抑制作用。方法采用荧光光谱法,研究不同浓度C085与Hsp90、NHsp90、MHsp90、CHsp90的相互作用以及293K、303K和310K下,C085与Hsp90的相互作用；实验选取280nm为激发波长,290-510 nm的波长范围内进行荧光光谱扫描。采用孔雀绿磷钼酸铵-无机磷检测法,研究 C085对 Hsp90-ATPase 活性的抑制。结果C085解离常数为（11．163±0．316）μmol · L-1；C085与Hsp90之间的主要作用力为静电作用力；C085与Hsp90的C端片段结合能力最强。当ATP为1 mmol · L-1时, C085作用于Hsp90的IC50值为6．04μmol · L-1。结论经过荧光光谱分析,可以确定C085与Hsp90的结合机制,且C085能抑制Hsp90-ATPase活性。Aim To estimate the affinity between C085 and Hsp90 and the inhibitory effects of C085 on the activity of Hsp90 ATPase. Methods The fluores-cence spectrum experiment was applied to examine the affinity between different C085 concentrations and Hsp90 , NHsp90 , MHsp90 , CHsp90; fluorescence in-tensities were recorded in the range of 290-510 nm at 293 K, 303 K and 310 K, respectively;a colorimetric assay for inorganic phosphate based on the formation of a phosphomolybdate complex and subsequent reaction with malachite green was used to examine the inhibitory effects of C085 on the activity of Hsp90 ATPase. Re-sults The dissociation constant KD value of C085 was （11. 163 ± 0. 316 ） μmol · L-1 . The interaction be-tween C085 and Hsp90 was driven mainly by electro-static interaction. C085 showed strongest affinity with CHsp90. When the concentration of ATP was 1 mmol· L-1 ,the inhibition of Hsp90 ATPase activity of C085 with the IC50 value was 6. 04μmol·L-1 . Conclusions The interaction mechanism between C085 and Hsp90 can be analyzed by fluorescence spectrum. C085 shows strong inhibition ATPase activity of Hsp90 .

关 键 词：C085 HSP90 ATPASE活性 荧光光谱法 相互作用 抑制作用 

分 类 号：R282.71[医药卫生—中药学] R284.1[医药卫生—中医学]