Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein
Background/Aims: Klotho, a protein mainly produced in the kidney and released into circulating blood, contributes to the negative regulation of 1,25(OH)2D3 formation and is thus a powerful regulator of mineral metabolism. As β-glucuronidase, alpha Klotho protein further regulates the stability of se...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Karger Publishers
2014-12-01
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| Series: | Kidney & Blood Pressure Research |
| Subjects: | |
| Online Access: | http://www.karger.com/Article/FullText/368472 |
| _version_ | 1818408253675012096 |
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| author | Ahmad Almilaji Sabina Honisch Guilai Liu Bernat Elvira Sumant Singh Ajay Zohreh Hosseinzadeh Musaab Ahmed Carlos Munoz Mentor Sopjani Florian Lang |
| author_facet | Ahmad Almilaji Sabina Honisch Guilai Liu Bernat Elvira Sumant Singh Ajay Zohreh Hosseinzadeh Musaab Ahmed Carlos Munoz Mentor Sopjani Florian Lang |
| author_sort | Ahmad Almilaji |
| collection | DOAJ |
| description | Background/Aims: Klotho, a protein mainly produced in the kidney and released into circulating blood, contributes to the negative regulation of 1,25(OH)2D3 formation and is thus a powerful regulator of mineral metabolism. As β-glucuronidase, alpha Klotho protein further regulates the stability of several carriers and channels in the plasma membrane and thus regulates channel and transporter activity. Accordingly, alpha Klotho protein participates in the regulation of diverse functions seemingly unrelated to mineral metabolism including lymphocyte function. The present study explored the impact of alpha Klotho protein on the voltage gated K+ channel Kv1.3. Methods: cRNA encoding Kv1.3 (KCNA3) was injected into Xenopus oocytes and depolarization induced outward current in Kv1.3 expressing Xenopus oocytes determined utilizing dual electrode voltage clamp. Experiments were performed without or with prior treatment with recombinant human Klotho protein (50 ng/ml, 24 hours) in the absence or presence of a β-glucuronidase inhibitor D-saccharic acid-1,4-lactone (DSAL, 10 µM). Moreover, the voltage gated K+ current was determined in Jcam lymphoma cells by whole cell patch clamp following 24 hours incubation without or with recombinant human Klotho protein (50 ng/ml, 24 hours). Kv1.3 protein abundance in Jcam cells was determined utilising fluorescent antibodies in flow cytometry. Results: In Kv1.3 expressing Xenopus oocytes the Kv1.3 currents and the protein abundance of Kv1.3 were both significantly enhanced after treatment with recombinant human Klotho protein (50 ng/ml, 24 hours), an effect reversed by presence of DSAL. Moreover, treatment with recombinant human Klotho protein increased Kv currents and Kv1.3 protein abundance in Jcam cells. Conclusion: Alpha Klotho protein enhances Kv1.3 channel abundance and Kv1.3 currents in the plasma membrane, an effect depending on its β-glucuronidase activity. |
| first_indexed | 2024-12-14T09:40:48Z |
| format | Article |
| id | doaj.art-a8f4fb2ed05641feb4957d448356af61 |
| institution | Directory Open Access Journal |
| issn | 1420-4096 1423-0143 |
| language | English |
| last_indexed | 2024-12-14T09:40:48Z |
| publishDate | 2014-12-01 |
| publisher | Karger Publishers |
| record_format | Article |
| series | Kidney & Blood Pressure Research |
| spelling | doaj.art-a8f4fb2ed05641feb4957d448356af612022-12-21T23:07:48ZengKarger PublishersKidney & Blood Pressure Research1420-40961423-01432014-12-0139660962210.1159/000368472368472Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho ProteinAhmad AlmilajiSabina HonischGuilai LiuBernat ElviraSumant Singh AjayZohreh HosseinzadehMusaab AhmedCarlos MunozMentor SopjaniFlorian LangBackground/Aims: Klotho, a protein mainly produced in the kidney and released into circulating blood, contributes to the negative regulation of 1,25(OH)2D3 formation and is thus a powerful regulator of mineral metabolism. As β-glucuronidase, alpha Klotho protein further regulates the stability of several carriers and channels in the plasma membrane and thus regulates channel and transporter activity. Accordingly, alpha Klotho protein participates in the regulation of diverse functions seemingly unrelated to mineral metabolism including lymphocyte function. The present study explored the impact of alpha Klotho protein on the voltage gated K+ channel Kv1.3. Methods: cRNA encoding Kv1.3 (KCNA3) was injected into Xenopus oocytes and depolarization induced outward current in Kv1.3 expressing Xenopus oocytes determined utilizing dual electrode voltage clamp. Experiments were performed without or with prior treatment with recombinant human Klotho protein (50 ng/ml, 24 hours) in the absence or presence of a β-glucuronidase inhibitor D-saccharic acid-1,4-lactone (DSAL, 10 µM). Moreover, the voltage gated K+ current was determined in Jcam lymphoma cells by whole cell patch clamp following 24 hours incubation without or with recombinant human Klotho protein (50 ng/ml, 24 hours). Kv1.3 protein abundance in Jcam cells was determined utilising fluorescent antibodies in flow cytometry. Results: In Kv1.3 expressing Xenopus oocytes the Kv1.3 currents and the protein abundance of Kv1.3 were both significantly enhanced after treatment with recombinant human Klotho protein (50 ng/ml, 24 hours), an effect reversed by presence of DSAL. Moreover, treatment with recombinant human Klotho protein increased Kv currents and Kv1.3 protein abundance in Jcam cells. Conclusion: Alpha Klotho protein enhances Kv1.3 channel abundance and Kv1.3 currents in the plasma membrane, an effect depending on its β-glucuronidase activity.http://www.karger.com/Article/FullText/368472Alpha Klotho proteinRecombinant human Klotho proteinKv1.3 channelXenopus oocytesJcam cells |
| spellingShingle | Ahmad Almilaji Sabina Honisch Guilai Liu Bernat Elvira Sumant Singh Ajay Zohreh Hosseinzadeh Musaab Ahmed Carlos Munoz Mentor Sopjani Florian Lang Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein Kidney & Blood Pressure Research Alpha Klotho protein Recombinant human Klotho protein Kv1.3 channel Xenopus oocytes Jcam cells |
| title | Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein |
| title_full | Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein |
| title_fullStr | Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein |
| title_full_unstemmed | Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein |
| title_short | Regulation of the Voltage Gated K+ Channel Kv1.3 by Recombinant Human Klotho Protein |
| title_sort | regulation of the voltage gated k channel kv1 3 by recombinant human klotho protein |
| topic | Alpha Klotho protein Recombinant human Klotho protein Kv1.3 channel Xenopus oocytes Jcam cells |
| url | http://www.karger.com/Article/FullText/368472 |
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