Design, synthesis and biological application of affinity-based small molecular probe for Hsp90 endoplasmic reticulum paralogue of Grp94
10.11665/j.issn.1000-5048.20190205
- VernacularTitle:热休克蛋白内质网亚型Grp94特异性荧光探针的设计、合成与应用
- Author:
Anping GUO
1
;
Fen JIANG
;
Xiaoli XU
;
Qidong YOU
;
Yuyan LI
Author Information
1. 中国药科大学江苏省药物分子设计与成药性优化重点实验室；药物化学系
- Publication Type:Journal Article
- Keywords:
heat shock proteins;
glucose-regulated protein 94;
synthesis;
fluorescence probe;
benzamide;
fluorescence polarization
- From:
Journal of China Pharmaceutical University
2019;50(2):161-167
- CountryChina
- Language:Chinese
-
Abstract:
Glucose-regulated protein 94(Grp94), an endoplasmic reticulum resident Hsp90 paralog, has a limited set of client proteins. Selective inhibition of Grp94 has emerged as a new direction for the development of drugs targeting the Hsp90 chaperone system. Now Grp94-Probe, an affinity-based probe of Grp94, was designed and synthesized based on DDO-5813, a most potent Grp94-selective inhibitor we found previously. Using fluorescence polarization(FP)assay and double staining assay with ER-Red in cells, we confirmed the binding of Grp94-Probe with ER Grp94. The FR results showed that the probe exhibited high affinity for Grp94N(EC50=117. 9 nmol/L)without exhibiting obvious Hsp90α inhibition, Moreover, as a fluorescence probe molecule, Grp94-Probe could better distinguish the inhibitory activity of compounds for Grp94N. The results of fluorescence analysis in cells showed that Grp94-Probe could co-stain with ER-Red in the endoplasmic reticulum, and the fluorescence did not decay rapidly with time after 4 h of staining, which further indicated the binding of Grp94-Probe with Grp94 in cells. This Grp94 selective probe can be further used for biology evaluation of Grp94 inhibitor and exploration of Grp94 biological functions.
