The Kv1.3 potassium channel is localized to the cis-Golgi and Kv1.6 is localized to the endoplasmic reticulum in rat astrocytes

Jing Zhu, Jenny Yan, William B. Thornhill

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

The functions of voltage-gated potassium (Kv) channels in neurons have been well defined, whereas their roles in glial cells are not fully understood. Kv1.1, Kv1.3 and Kv1.6 are endogenously expressed in C6 astrocytoma cells, but their trafficking and subcellular localization have not been well studied. In C6 cells, Kv1.1 was localized to the cell surface, Kv1.3 was predominantly localized in the cis-Golgi, and Kv1.6 was enriched in the endoplasmic reticulum. Disruption of the Golgi stacks with brefeldin A treatment redirected Kv1.3 to the endoplasmic reticulum, further confirming that Kv1.3 was localized in the Golgi. Denaturing and reducing immunoblot analysis identified an expected Kv1.3 monomer and an unexpected Kv1.3 dimer/aggregate. These two forms had different protein half-lives: that of the monomer form T1/2 was 5.1 h, whereas the dimer/aggregate form was stable over the 8-h measurement period. The Kv1.3 dimer/aggregate form on immunoblots appeared to be correlated with its Golgi retention, based on examination with several cell types that expressed Kv1.3. Glycosidase treatment showed that Kv1.3 contained complex-type N-glycans terminated with sialic acids, suggesting that Kv1.3 had traveled to the trans-Golgi network for sialylation before it was recycled to the cis-Golgi for retention. Inhibition of N-glycosylation did not affect Kv1.3 localization, indicating that N-glycans did not play a role in its Golgi retention. Thus, Kv1.3 appears to be distributed to the cis-Golgi membrane of rat astrocytes in a similar way as a Golgi resident protein, and this unusual distribution appears to be correlated with its SDS/2-mercaptoethanol-resistant dimer/aggregate forms on immunoblots.

Original languageEnglish (US)
Pages (from-to)3433-3445
Number of pages13
JournalFEBS Journal
Volume281
Issue number15
DOIs
StatePublished - Aug 2014
Externally publishedYes

Keywords

  • Golgi
  • astrocyte
  • endoplasmic reticulum
  • potassium channel
  • subcellular localization

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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