Topology of transmembrane channel-like gene 1 protein

Valentina Labay, Rachel M. Weichert, Tomoko Makishima, Andrew J. Griffith

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Mutations of transmembrane channel-like gene 1 (TMC1) cause hearing loss in humans and mice. TMC1 is the founding member of a family of genes encoding proteins of unknown function that are predicted to contain multiple transmembrane domains. The goal of our study was to define the topology of mouse TMC1 expressed heterologously in tissue culture cells. TMC1 was retained in the endoplasmic reticulum (ER) membrane of five tissue culture cell lines that we tested. We used anti-TMC1 and anti-HA antibodies to probe the topologic orientation of three native epitopes and seven HA epitope tags along full-length TMC1 after selective or complete permeabilization of transfected cells with digitonin or Triton X-100, respectively. TMC1 was present within the ER as an integral membrane protein containing six transmembrane domains and cytosolic N- and C-termini. There is a large cytoplasmic loop, between the fourth and fifth transmembrane domains, with two highly conserved hydrophobic regions that might associate with or penetrate, but do not span, the plasma membrane. Our study is the first to demonstrate that TMC1 is a transmembrane protein. The topologic organization revealed by this study shares some features with that of the shaker-TRP superfamily of ion channels.

Original languageEnglish (US)
Pages (from-to)8592-8598
Number of pages7
JournalBiochemistry
Volume49
Issue number39
DOIs
StatePublished - Oct 5 2010
Externally publishedYes

Fingerprint

Genes
Topology
Proteins
Tissue culture
Endoplasmic Reticulum
Epitopes
Digitonin
Gene encoding
Octoxynol
Audition
Cell membranes
Ion Channels
Hearing Loss
Anti-Idiotypic Antibodies
Membrane Proteins
Cell Culture Techniques
Cell Membrane
Organizations
Membranes
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Labay, V., Weichert, R. M., Makishima, T., & Griffith, A. J. (2010). Topology of transmembrane channel-like gene 1 protein. Biochemistry, 49(39), 8592-8598. https://doi.org/10.1021/bi1004377

Topology of transmembrane channel-like gene 1 protein. / Labay, Valentina; Weichert, Rachel M.; Makishima, Tomoko; Griffith, Andrew J.

In: Biochemistry, Vol. 49, No. 39, 05.10.2010, p. 8592-8598.

Research output: Contribution to journalArticle

Labay, V, Weichert, RM, Makishima, T & Griffith, AJ 2010, 'Topology of transmembrane channel-like gene 1 protein', Biochemistry, vol. 49, no. 39, pp. 8592-8598. https://doi.org/10.1021/bi1004377
Labay, Valentina ; Weichert, Rachel M. ; Makishima, Tomoko ; Griffith, Andrew J. / Topology of transmembrane channel-like gene 1 protein. In: Biochemistry. 2010 ; Vol. 49, No. 39. pp. 8592-8598.
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