Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes
In the present study, we synthesized and, structurally and functionally characterized a novel α4/7-conotoxin Mr1.7 (PECCTHPACHVSHPELC-NH2), which was previously identified by cDNA libraries from Conus marmoreus in our lab. The NMR solution structure showed that Mr1.7 contained a 310-helix from resid...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2015-05-01
|
| Series: | Marine Drugs |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1660-3397/13/6/3259 |
| _version_ | 1828347855011577856 |
|---|---|
| author | Shuo Wang Cong Zhao Zhuguo Liu Xuesong Wang Na Liu Weihong Du Qiuyun Dai |
| author_facet | Shuo Wang Cong Zhao Zhuguo Liu Xuesong Wang Na Liu Weihong Du Qiuyun Dai |
| author_sort | Shuo Wang |
| collection | DOAJ |
| description | In the present study, we synthesized and, structurally and functionally characterized a novel α4/7-conotoxin Mr1.7 (PECCTHPACHVSHPELC-NH2), which was previously identified by cDNA libraries from Conus marmoreus in our lab. The NMR solution structure showed that Mr1.7 contained a 310-helix from residues Pro7 to His10 and a type I β-turn from residues Pro14 to Cys17. Electrophysiological results showed that Mr1.7 selectively inhibited the α3β2, α9α10 and α6/α3β2β3 neuronal nicotinic acetylcholine receptors (nAChRs) with an IC50 of 53.1 nM, 185.7 nM and 284.2 nM, respectively, but showed no inhibitory activity on other nAChR subtypes. Further structure-activity studies of Mr1.7 demonstrated that the PE residues at the N-terminal sequence of Mr1.7 were important for modulating its selectivity, and the replacement of Glu2 by Ala resulted in a significant increase in potency and selectivity to the α3β2 nAChR. Furthermore, the substitution of Ser12 with Asn in the loop2 significantly increased the binding of Mr1.7 to α3β2, α3β4, α2β4 and α7 nAChR subtypes. Taken together, this work expanded our knowledge of selectivity and provided a new way to improve the potency and selectivity of inhibitors for nAChR subtypes. |
| first_indexed | 2024-04-14T00:46:20Z |
| format | Article |
| id | doaj.art-a149808813ce4a259c535aff6ab86cf8 |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-14T00:46:20Z |
| publishDate | 2015-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-a149808813ce4a259c535aff6ab86cf82022-12-22T02:22:01ZengMDPI AGMarine Drugs1660-33972015-05-011363259327510.3390/md13063259md13063259Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 SubtypesShuo Wang0Cong Zhao1Zhuguo Liu2Xuesong Wang3Na Liu4Weihong Du5Qiuyun Dai6Beijing Institute of Biotechnology, Beijing 100071, ChinaDepartment of Chemistry, Renmin University of China, Beijing 100872, ChinaBeijing Institute of Biotechnology, Beijing 100071, ChinaDepartment of Chemistry, Renmin University of China, Beijing 100872, ChinaBeijing Institute of Biotechnology, Beijing 100071, ChinaDepartment of Chemistry, Renmin University of China, Beijing 100872, ChinaBeijing Institute of Biotechnology, Beijing 100071, ChinaIn the present study, we synthesized and, structurally and functionally characterized a novel α4/7-conotoxin Mr1.7 (PECCTHPACHVSHPELC-NH2), which was previously identified by cDNA libraries from Conus marmoreus in our lab. The NMR solution structure showed that Mr1.7 contained a 310-helix from residues Pro7 to His10 and a type I β-turn from residues Pro14 to Cys17. Electrophysiological results showed that Mr1.7 selectively inhibited the α3β2, α9α10 and α6/α3β2β3 neuronal nicotinic acetylcholine receptors (nAChRs) with an IC50 of 53.1 nM, 185.7 nM and 284.2 nM, respectively, but showed no inhibitory activity on other nAChR subtypes. Further structure-activity studies of Mr1.7 demonstrated that the PE residues at the N-terminal sequence of Mr1.7 were important for modulating its selectivity, and the replacement of Glu2 by Ala resulted in a significant increase in potency and selectivity to the α3β2 nAChR. Furthermore, the substitution of Ser12 with Asn in the loop2 significantly increased the binding of Mr1.7 to α3β2, α3β4, α2β4 and α7 nAChR subtypes. Taken together, this work expanded our knowledge of selectivity and provided a new way to improve the potency and selectivity of inhibitors for nAChR subtypes.http://www.mdpi.com/1660-3397/13/6/3259neuronal nicotinic acetylcholine receptorα-conotoxin Mr1.7Conus marmoreusselectivityN-terminal sequencestructure-activity relationship |
| spellingShingle | Shuo Wang Cong Zhao Zhuguo Liu Xuesong Wang Na Liu Weihong Du Qiuyun Dai Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes Marine Drugs neuronal nicotinic acetylcholine receptor α-conotoxin Mr1.7 Conus marmoreus selectivity N-terminal sequence structure-activity relationship |
| title | Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes |
| title_full | Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes |
| title_fullStr | Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes |
| title_full_unstemmed | Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes |
| title_short | Structural and Functional Characterization of a Novel α-Conotoxin Mr1.7 from Conus marmoreus Targeting Neuronal nAChR α3β2, α9α10 and α6/α3β2β3 Subtypes |
| title_sort | structural and functional characterization of a novel α conotoxin mr1 7 from conus marmoreus targeting neuronal nachr α3β2 α9α10 and α6 α3β2β3 subtypes |
| topic | neuronal nicotinic acetylcholine receptor α-conotoxin Mr1.7 Conus marmoreus selectivity N-terminal sequence structure-activity relationship |
| url | http://www.mdpi.com/1660-3397/13/6/3259 |
| work_keys_str_mv | AT shuowang structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes AT congzhao structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes AT zhuguoliu structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes AT xuesongwang structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes AT naliu structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes AT weihongdu structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes AT qiuyundai structuralandfunctionalcharacterizationofanovelaconotoxinmr17fromconusmarmoreustargetingneuronalnachra3b2a9a10anda6a3b2b3subtypes |