Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel
The hERG (human ether à go-go related gene) potassium channel is a voltage-gated potassium channel playing important roles in the heart by controlling the rapid delayed rectifier potassium current. The hERG protein contains a voltage-sensor domain (VSD) that is important for sensing voltage changes...
| Main Authors: | , , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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2013
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| Online Access: | https://hdl.handle.net/10356/99423 http://hdl.handle.net/10220/10951 |
| _version_ | 1826112446736105472 |
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| author | Ng, Hui Qi Kim, Young Mee Huang, Qiwei Gayen, Shovanlal Yildiz, Ahu Arslan Yoon, Ho Sup Sinner, Eva-Kathrin Kang, CongBao |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Ng, Hui Qi Kim, Young Mee Huang, Qiwei Gayen, Shovanlal Yildiz, Ahu Arslan Yoon, Ho Sup Sinner, Eva-Kathrin Kang, CongBao |
| author_sort | Ng, Hui Qi |
| collection | NTU |
| description | The hERG (human ether à go-go related gene) potassium channel is a voltage-gated potassium channel playing important roles in the heart by controlling the rapid delayed rectifier potassium current. The hERG protein contains a voltage-sensor domain (VSD) that is important for sensing voltage changes across the membrane. Mutations in this domain contribute to serious heart diseases. To study the structure of the VSD, it was over-expressed in Escherichia coli and purified into detergent micelles. Lyso-myristoyl phosphatidylglycerol (LMPG) was shown to be a suitable detergent for VSD purification and folding. Secondary structural analysis using circular dichroism (CD) spectroscopy indicated that the purified VSD in LMPG micelles adopted α-helical structures. Purified VSD in LMPG micelles produced dispersed cross-peaks in a 15N-HSQC spectrum. Backbone resonance assignments for residues from transmembrane segments S3 and S4 of VSD also confirmed the presence of α-helical structures in this domain. Our results demonstrated that structure of VSD can be investigated using NMR spectroscopy. |
| first_indexed | 2024-10-01T03:07:09Z |
| format | Journal Article |
| id | ntu-10356/99423 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:07:09Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | ntu-10356/994232020-03-07T12:18:19Z Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel Ng, Hui Qi Kim, Young Mee Huang, Qiwei Gayen, Shovanlal Yildiz, Ahu Arslan Yoon, Ho Sup Sinner, Eva-Kathrin Kang, CongBao School of Biological Sciences DRNTU::Science::Biological sciences The hERG (human ether à go-go related gene) potassium channel is a voltage-gated potassium channel playing important roles in the heart by controlling the rapid delayed rectifier potassium current. The hERG protein contains a voltage-sensor domain (VSD) that is important for sensing voltage changes across the membrane. Mutations in this domain contribute to serious heart diseases. To study the structure of the VSD, it was over-expressed in Escherichia coli and purified into detergent micelles. Lyso-myristoyl phosphatidylglycerol (LMPG) was shown to be a suitable detergent for VSD purification and folding. Secondary structural analysis using circular dichroism (CD) spectroscopy indicated that the purified VSD in LMPG micelles adopted α-helical structures. Purified VSD in LMPG micelles produced dispersed cross-peaks in a 15N-HSQC spectrum. Backbone resonance assignments for residues from transmembrane segments S3 and S4 of VSD also confirmed the presence of α-helical structures in this domain. Our results demonstrated that structure of VSD can be investigated using NMR spectroscopy. 2013-07-04T04:17:47Z 2019-12-06T20:07:02Z 2013-07-04T04:17:47Z 2019-12-06T20:07:02Z 2012 2012 Journal Article Ng, H. Q., Kim, Y. M., Huang, Q., Gayen, S., Yildiz, A. A., Yoon, H. S. (2012). Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel. Protein Expression and Purification, 86(2), 98–104. 1046-5928 https://hdl.handle.net/10356/99423 http://hdl.handle.net/10220/10951 10.1016/j.pep.2012.09.003 en Protein expression and purification © 2012 Elsevier Inc. |
| spellingShingle | DRNTU::Science::Biological sciences Ng, Hui Qi Kim, Young Mee Huang, Qiwei Gayen, Shovanlal Yildiz, Ahu Arslan Yoon, Ho Sup Sinner, Eva-Kathrin Kang, CongBao Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel |
| title | Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel |
| title_full | Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel |
| title_fullStr | Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel |
| title_full_unstemmed | Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel |
| title_short | Purification and structural characterization of the voltage-sensor domain of the hERG potassium channel |
| title_sort | purification and structural characterization of the voltage sensor domain of the herg potassium channel |
| topic | DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/99423 http://hdl.handle.net/10220/10951 |
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