μl Na+ channels expressed transiently in human embryonic kidney cells: Biochemical and biophysical properties

Chinweike Ukomadu; Jiuying Zhou; Fred J. Sigworth; William S. Agnew

doi:10.1016/0896-6273(92)90088-U

μl Na⁺ channels expressed transiently in human embryonic kidney cells: Biochemical and biophysical properties

Chinweike Ukomadu, Jiuying Zhou, Fred J. Sigworth, William S. Agnew

Research output: Contribution to journal › Article › peer-review

Abstract

We describe the transient expression of the rat skeletal muscle μl Na⁺ channel in human embryonic kidney (HEK 293) cells. Functional channels appear at a density of ∼30 in a 10 μm² patch, comparable to those of native excitable cells. Unlike pl currents in oocytes, inactivation gating is predominantly (∼97%) fast, although clear evidence is provided for noninactivating gating modes, which have been linked to anomalous behavior in the inherited disorder hyperkalemic periodic paralysis. Sequence-specific antibodies detect a ∼230 kd glycopeptide. The majority of molecules acquire only neutral oligosaccharides and are retained within the cell. Electrophoretic mobility on SDS gels suggests the molecules may acquire covalently attached lipid. The channel is readily phosphorylated by activation of the protein kinase A and protein kinase C second messenger pathways.

Original language	English (US)
Pages (from-to)	663-676
Number of pages	14
Journal	Neuron
Volume	8
Issue number	4
DOIs	https://doi.org/10.1016/0896-6273(92)90088-U
State	Published - Apr 1992
Externally published	Yes

ASJC Scopus subject areas

Neuroscience(all)

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title = "μl Na+ channels expressed transiently in human embryonic kidney cells: Biochemical and biophysical properties",

abstract = "We describe the transient expression of the rat skeletal muscle μl Na+ channel in human embryonic kidney (HEK 293) cells. Functional channels appear at a density of ∼30 in a 10 μm2 patch, comparable to those of native excitable cells. Unlike pl currents in oocytes, inactivation gating is predominantly (∼97%) fast, although clear evidence is provided for noninactivating gating modes, which have been linked to anomalous behavior in the inherited disorder hyperkalemic periodic paralysis. Sequence-specific antibodies detect a ∼230 kd glycopeptide. The majority of molecules acquire only neutral oligosaccharides and are retained within the cell. Electrophoretic mobility on SDS gels suggests the molecules may acquire covalently attached lipid. The channel is readily phosphorylated by activation of the protein kinase A and protein kinase C second messenger pathways.",

author = "Chinweike Ukomadu and Jiuying Zhou and Sigworth, {Fred J.} and Agnew, {William S.}",

note = "Funding Information: We thank Dr. R. Kopito for providing the vector RBG4 and the HEK 293 cells. We also thank Mike Maceyka, Jerry Potts, Jim Trimmer, Sunil Hingorani, Megan Stephan, Doris Lin, and Steve Sine for helpful discussions. This work was supported by National Institutes of Health grants HL 38156 and NS 17928 and a grant from the Muscular Dystrophy Association and from the National Multiple Sclerosis Society. C. U. is a Howard Hughes predoctoral fellow. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 USC Section 1734 solely to indicate this fact. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.",

year = "1992",

month = apr,

doi = "10.1016/0896-6273(92)90088-U",

language = "English (US)",

volume = "8",

pages = "663--676",

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AU - Ukomadu, Chinweike

AU - Zhou, Jiuying

AU - Sigworth, Fred J.

AU - Agnew, William S.

N1 - Funding Information: We thank Dr. R. Kopito for providing the vector RBG4 and the HEK 293 cells. We also thank Mike Maceyka, Jerry Potts, Jim Trimmer, Sunil Hingorani, Megan Stephan, Doris Lin, and Steve Sine for helpful discussions. This work was supported by National Institutes of Health grants HL 38156 and NS 17928 and a grant from the Muscular Dystrophy Association and from the National Multiple Sclerosis Society. C. U. is a Howard Hughes predoctoral fellow. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 USC Section 1734 solely to indicate this fact. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.

PY - 1992/4

Y1 - 1992/4

N2 - We describe the transient expression of the rat skeletal muscle μl Na+ channel in human embryonic kidney (HEK 293) cells. Functional channels appear at a density of ∼30 in a 10 μm2 patch, comparable to those of native excitable cells. Unlike pl currents in oocytes, inactivation gating is predominantly (∼97%) fast, although clear evidence is provided for noninactivating gating modes, which have been linked to anomalous behavior in the inherited disorder hyperkalemic periodic paralysis. Sequence-specific antibodies detect a ∼230 kd glycopeptide. The majority of molecules acquire only neutral oligosaccharides and are retained within the cell. Electrophoretic mobility on SDS gels suggests the molecules may acquire covalently attached lipid. The channel is readily phosphorylated by activation of the protein kinase A and protein kinase C second messenger pathways.

AB - We describe the transient expression of the rat skeletal muscle μl Na+ channel in human embryonic kidney (HEK 293) cells. Functional channels appear at a density of ∼30 in a 10 μm2 patch, comparable to those of native excitable cells. Unlike pl currents in oocytes, inactivation gating is predominantly (∼97%) fast, although clear evidence is provided for noninactivating gating modes, which have been linked to anomalous behavior in the inherited disorder hyperkalemic periodic paralysis. Sequence-specific antibodies detect a ∼230 kd glycopeptide. The majority of molecules acquire only neutral oligosaccharides and are retained within the cell. Electrophoretic mobility on SDS gels suggests the molecules may acquire covalently attached lipid. The channel is readily phosphorylated by activation of the protein kinase A and protein kinase C second messenger pathways.

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μl Na⁺ channels expressed transiently in human embryonic kidney cells: Biochemical and biophysical properties

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