Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels
Abstract Background KCNH family of potassium channels is responsible for diverse physiological functions ranging from the regulation of neuronal excitability and cardiac contraction to the regulation of cancer progression. KCNH channels contain a Per-Arn-Sim (PAS) domain in their N-terminal and cycl...
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| Format: | Article |
| Language: | English |
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BMC
2019-07-01
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| Series: | BMC Pharmacology and Toxicology |
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| Online Access: | http://link.springer.com/article/10.1186/s40360-019-0324-8 |
| _version_ | 1798045824953876480 |
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| author | Ze-Jun Wang Purushottam B. Tiwari Aykut Üren Tinatin I. Brelidze |
| author_facet | Ze-Jun Wang Purushottam B. Tiwari Aykut Üren Tinatin I. Brelidze |
| author_sort | Ze-Jun Wang |
| collection | DOAJ |
| description | Abstract Background KCNH family of potassium channels is responsible for diverse physiological functions ranging from the regulation of neuronal excitability and cardiac contraction to the regulation of cancer progression. KCNH channels contain a Per-Arn-Sim (PAS) domain in their N-terminal and cyclic nucleotide-binding homology (CNBH) domain in their C-terminal regions. These intracellular domains shape the function of KCNH channels and are important targets for drug development. Methods Here we describe a surface plasmon resonance (SPR)-based screening method aimed in identifying small molecule binders of PAS and CNBH domains for three KCNH channel subfamilies: ether-à-go-go (EAG), EAG-related gene (ERG), and EAG-like K+ (ELK). The method involves purification of the PAS and CNBH domains, immobilization of the purified domains on the SPR senor chip and screening small molecules in a chemical library for binding to the immobilized domains using changes in the SPR response as a reporter of the binding. The advantages of this method include low quantity of purified PAS and CNBH domains necessary for the implementation of the screen, direct assessment of the small molecule binding to the PAS and CNBH domains and easiness of assessing KCNH subfamily specificity of the small molecule binders. Results Using the SPR-based method we screened the Spectrum Collection Library of 2560 compounds against the PAS and CNBH domains of the three KCNH channel subfamilies and identified a pool of small molecules that bind to the PAS or CNBH domains. To further evaluate the effectiveness of the screen we tested the functional effect of one of the identified mEAG PAS domain specific small molecule binders on currents recorded from EAG channels. Undecylenic acid inhibited currents recorded from EAG channels in a concentration-dependent manner with IC50 of ~ 1 μM. Conclusion Our results show that the SPR-based method is well suited for identifying small molecule binders of KCNH channels and can facilitate drug discovery for other ion channels as well. |
| first_indexed | 2024-04-11T23:27:21Z |
| format | Article |
| id | doaj.art-30a3f286c17a42cea8c681515b173005 |
| institution | Directory Open Access Journal |
| issn | 2050-6511 |
| language | English |
| last_indexed | 2024-04-11T23:27:21Z |
| publishDate | 2019-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Pharmacology and Toxicology |
| spelling | doaj.art-30a3f286c17a42cea8c681515b1730052022-12-22T03:57:16ZengBMCBMC Pharmacology and Toxicology2050-65112019-07-0120111110.1186/s40360-019-0324-8Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channelsZe-Jun Wang0Purushottam B. Tiwari1Aykut Üren2Tinatin I. Brelidze3Department of Pharmacology and Physiology, Georgetown University Medical CenterDepartment of Oncology, Georgetown University Medical CenterDepartment of Oncology, Georgetown University Medical CenterDepartment of Pharmacology and Physiology, Georgetown University Medical CenterAbstract Background KCNH family of potassium channels is responsible for diverse physiological functions ranging from the regulation of neuronal excitability and cardiac contraction to the regulation of cancer progression. KCNH channels contain a Per-Arn-Sim (PAS) domain in their N-terminal and cyclic nucleotide-binding homology (CNBH) domain in their C-terminal regions. These intracellular domains shape the function of KCNH channels and are important targets for drug development. Methods Here we describe a surface plasmon resonance (SPR)-based screening method aimed in identifying small molecule binders of PAS and CNBH domains for three KCNH channel subfamilies: ether-à-go-go (EAG), EAG-related gene (ERG), and EAG-like K+ (ELK). The method involves purification of the PAS and CNBH domains, immobilization of the purified domains on the SPR senor chip and screening small molecules in a chemical library for binding to the immobilized domains using changes in the SPR response as a reporter of the binding. The advantages of this method include low quantity of purified PAS and CNBH domains necessary for the implementation of the screen, direct assessment of the small molecule binding to the PAS and CNBH domains and easiness of assessing KCNH subfamily specificity of the small molecule binders. Results Using the SPR-based method we screened the Spectrum Collection Library of 2560 compounds against the PAS and CNBH domains of the three KCNH channel subfamilies and identified a pool of small molecules that bind to the PAS or CNBH domains. To further evaluate the effectiveness of the screen we tested the functional effect of one of the identified mEAG PAS domain specific small molecule binders on currents recorded from EAG channels. Undecylenic acid inhibited currents recorded from EAG channels in a concentration-dependent manner with IC50 of ~ 1 μM. Conclusion Our results show that the SPR-based method is well suited for identifying small molecule binders of KCNH channels and can facilitate drug discovery for other ion channels as well.http://link.springer.com/article/10.1186/s40360-019-0324-8PAS domainCNBH domainDrug screening |
| spellingShingle | Ze-Jun Wang Purushottam B. Tiwari Aykut Üren Tinatin I. Brelidze Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels BMC Pharmacology and Toxicology PAS domain CNBH domain Drug screening |
| title | Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels |
| title_full | Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels |
| title_fullStr | Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels |
| title_full_unstemmed | Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels |
| title_short | Identification of undecylenic acid as EAG channel inhibitor using surface plasmon resonance-based screen of KCNH channels |
| title_sort | identification of undecylenic acid as eag channel inhibitor using surface plasmon resonance based screen of kcnh channels |
| topic | PAS domain CNBH domain Drug screening |
| url | http://link.springer.com/article/10.1186/s40360-019-0324-8 |
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