Localization of the nucleic acid channel regulatory subunit, cytosolic malate dehydrogenase

Basil Hanss, Edgar Leal-Pinto, Avelino Teixeira, Baohuong Tran, Chun Hui Lee, Scott C. Henderson, Paul E. Klotman

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

NACh is a nucleic acid-conducting channel found in apical membrane of rat kidney proximal tubules. It is a heteromultimeric complex consisting of at least two proteins: a 45-kDa pore-forming subunit and a 36-kDa regulatory subunit. The regulatory subunit confers ion selectivity and influences gating kinetics. The regulatory subunit has been identified as cytosolic malate dehydrogenase (cMDH). cMDH is described in the literature as a soluble protein that is not associated with plasma membrane. Yet a role for cMDH as the regulatory subunit of NACh requires that it be present at the plasma membrane. To resolve this conflict, studies were initiated to determine whether cMDH could be found at the plasma membrane. Before performing localization studies, a suitable model system that expressed NACh was identified. A channel was identified in LLC-PK1 cells, a line derived from pig proximal tubule, that is selective for nucleic acid and has a conductance of approximately 10 pS. It exhibits dose-dependent blockade by heparan sulfate or l-malate. These characteristics are similar to what has been reported for NACh from rat kidney and indicate that NACh is present in LLC-PK1 cells. LLC-PK 1 cells were therefore used as a model system for immunolocalization of cMDH. Both immunofluorescence and immunoelectron microscopy demonstrated cMDH at the plasma membrane of LLC-PK1 cells. This finding supports prior functional data that describe a role for cMDH as the regulatory subunit of NACh.

Original languageEnglish
Pages (from-to)1-8
Number of pages8
JournalJournal of Membrane Biology
Volume226
Issue number1-3
DOIs
StatePublished - Dec 2008

Keywords

  • Artifical planar membranes
  • Electrophysiology
  • Epithelial ion transport
  • Ion channels
  • Ion transport by epithelia
  • Membrane biology/immuno chemistry
  • Membrane transport
  • Planar bilayers
  • Renal physiology

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