Importance of glycosylation on function of a potassium channel in neuroblastoma cells.
The Kv3.1 glycoprotein, a voltage-gated potassium channel, is expressed throughout the central nervous system. The role of N-glycans attached to the Kv3.1 glycoprotein on conducting and non-conducting functions of the Kv3.1 channel are quite limiting. Glycosylated (wild type), partially glycosylated...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3082577?pdf=render |
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| author | M K Hall Tara A Cartwright Christa M Fleming Ruth A Schwalbe |
| author_facet | M K Hall Tara A Cartwright Christa M Fleming Ruth A Schwalbe |
| author_sort | M K Hall |
| collection | DOAJ |
| description | The Kv3.1 glycoprotein, a voltage-gated potassium channel, is expressed throughout the central nervous system. The role of N-glycans attached to the Kv3.1 glycoprotein on conducting and non-conducting functions of the Kv3.1 channel are quite limiting. Glycosylated (wild type), partially glycosylated (N220Q and N229Q), and unglycosylated (N220Q/N229Q) Kv3.1 proteins were expressed and characterized in a cultured neuronal-derived cell model, B35 neuroblastoma cells. Western blots, whole cell current recordings, and wound healing assays were employed to provide evidence that the conducting and non-conducting properties of the Kv3.1 channel were modified by N-glycans of the Kv3.1 glycoprotein. Electrophoretic migration of the various Kv3.1 proteins treated with PNGase F and neuraminidase verified that the glycosylation sites were occupied and that the N-glycans could be sialylated, respectively. The unglycosylated channel favored a different whole cell current pattern than the glycoform. Further the outward ionic currents of the unglycosylated channel had slower activation and deactivation rates than those of the glycosylated Kv3.1 channel. These kinetic parameters of the partially glycosylated Kv3.1 channels were also slowed. B35 cells expressing glycosylated Kv3.1 protein migrated faster than those expressing partially glycosylated and much faster than those expressing the unglycosylated Kv3.1 protein. These results have demonstrated that N-glycans of the Kv3.1 glycoprotein enhance outward ionic current kinetics, and neuronal migration. It is speculated that physiological changes which lead to a reduction in N-glycan attachment to proteins will alter the functions of the Kv3.1 channel. |
| first_indexed | 2024-12-14T17:33:18Z |
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| id | doaj.art-e64b2147193f49b0997599871d9c43ac |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T17:33:18Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-e64b2147193f49b0997599871d9c43ac2022-12-21T22:53:03ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0164e1931710.1371/journal.pone.0019317Importance of glycosylation on function of a potassium channel in neuroblastoma cells.M K HallTara A CartwrightChrista M FlemingRuth A SchwalbeThe Kv3.1 glycoprotein, a voltage-gated potassium channel, is expressed throughout the central nervous system. The role of N-glycans attached to the Kv3.1 glycoprotein on conducting and non-conducting functions of the Kv3.1 channel are quite limiting. Glycosylated (wild type), partially glycosylated (N220Q and N229Q), and unglycosylated (N220Q/N229Q) Kv3.1 proteins were expressed and characterized in a cultured neuronal-derived cell model, B35 neuroblastoma cells. Western blots, whole cell current recordings, and wound healing assays were employed to provide evidence that the conducting and non-conducting properties of the Kv3.1 channel were modified by N-glycans of the Kv3.1 glycoprotein. Electrophoretic migration of the various Kv3.1 proteins treated with PNGase F and neuraminidase verified that the glycosylation sites were occupied and that the N-glycans could be sialylated, respectively. The unglycosylated channel favored a different whole cell current pattern than the glycoform. Further the outward ionic currents of the unglycosylated channel had slower activation and deactivation rates than those of the glycosylated Kv3.1 channel. These kinetic parameters of the partially glycosylated Kv3.1 channels were also slowed. B35 cells expressing glycosylated Kv3.1 protein migrated faster than those expressing partially glycosylated and much faster than those expressing the unglycosylated Kv3.1 protein. These results have demonstrated that N-glycans of the Kv3.1 glycoprotein enhance outward ionic current kinetics, and neuronal migration. It is speculated that physiological changes which lead to a reduction in N-glycan attachment to proteins will alter the functions of the Kv3.1 channel.http://europepmc.org/articles/PMC3082577?pdf=render |
| spellingShingle | M K Hall Tara A Cartwright Christa M Fleming Ruth A Schwalbe Importance of glycosylation on function of a potassium channel in neuroblastoma cells. PLoS ONE |
| title | Importance of glycosylation on function of a potassium channel in neuroblastoma cells. |
| title_full | Importance of glycosylation on function of a potassium channel in neuroblastoma cells. |
| title_fullStr | Importance of glycosylation on function of a potassium channel in neuroblastoma cells. |
| title_full_unstemmed | Importance of glycosylation on function of a potassium channel in neuroblastoma cells. |
| title_short | Importance of glycosylation on function of a potassium channel in neuroblastoma cells. |
| title_sort | importance of glycosylation on function of a potassium channel in neuroblastoma cells |
| url | http://europepmc.org/articles/PMC3082577?pdf=render |
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