Design, synthesis, and activity study of α-conotoxin A10LPnIA fluorescent probe
10.16438/j.0513-4870.2021-0528
- VernacularTitle:α-芋螺毒素[A10L]PnIA荧光探针的设计合成及活性研究
- Author:
Yao TAN
1
;
Yi-shuai YANG
1
;
Zhao-li CHU
1
;
Dong-ting ZHANGSUN
1
;
Xiao-peng ZHU
1
,
2
;
Su-lan LUO
1
,
2
Author Information
1. Key Laboratory of Tropical Biological Resources of Ministry of Education, School of Life and Pharmaceutical Sciences, Hainan University, Haikou 570228, China
2. Medical School, Guangxi University, Nanning 530004, China
- Publication Type:Research Article
- Keywords:
italic>α7 nicotinic acetylcholine receptor;
italic>α-conotoxin [A10L]PnIA;
fluorescent probe;
electrophysiological activity
- From:
Acta Pharmaceutica Sinica
2021;56(8):2252-2259
- CountryChina
- Language:Chinese
-
Abstract:
italic>α7 nicotinic acetylcholine receptor (nAChR) is widely distributed in the central and peripheral nervous systems, and is closely related to a variety of neurological diseases and inflammation response. α-Conotoxin [A10L]PnIA, as an antagonist targeting α7 nAChR, plays an important role in studying the physiological and pathological processes involved in α7 nAChR. [A10L]PnIA was labeled with fluorescein 5-carboxytetramethylrhodamine, and the active peptide ([A10L]PnIA-F) was obtained by a two-step oxidative folding procedure in vitro. The Xenopus oocyte expression system and the two-electrode voltage clamp technique were used to identify the potency of [A10L]PnIA-F fluorescent peptide, and its cytotoxicity was detected by mouse macrophages and CCK8 method. The molecular weight of [A10L]PnIA-F fluorescent peptide was identified by mass spectrometry as 2 077.28 Da, which was consistent with the theoretical value. Electrophysiological determination of its half-maximal inhibitory concentration (IC50) for α7 nAChR is 17.32 nmol·L-1, which is consistent with [A10L]PnIA (IC50, 13.84 nmol·L-1). The cytotoxicity test results showed that within the concentration range of 5 nmol·L-1 to 10 μmol·L-1, there was no significant inhibition on the growth of mouse macrophages. The results showed that the α-conotoxin fluorescent probe [A10L]PnIA could provide pharmacological tools for the research of α7 nAChR-related neurophysiological and pathological mechanisms.
