Functional dissection of transmembrane domains of human TAP-like (ABCB9)

Biochem Biophys Res Commun. 2008 Dec 19;377(3):847-51. doi: 10.1016/j.bbrc.2008.10.078. Epub 2008 Oct 24.

Abstract

An ABC transporter, TAP-Like (TAPL), was dissected into its amino-terminal transmembrane domain and the following core domain. When these domains were transiently expressed as tagged proteins with a His6- or Myc-epitope tag, the amino-terminal ones (Met(1)-Lys(182)) could not associate with each other, or with the full-length transporter (Met(1)-Ala(766)). However, both the core domain (Arg(141)-Ala(766)) and full-length protein mutually interacted. The amino-terminal domain (Met(1)-Arg(141)) as well as the full-length transporter fused with fluorescent protein GFP was sorted to lysosomal membranes upon their stable expression, as visualized by means of fluorescent microscopy, while the core domain (Arg(141)-Ala(766)) was broadly distributed in the intra-cellular membranes. These results suggest that the sorting signal for lysosomes is present within the amino-terminal transmembrane domain (Met(1)-Arg(141)) of the TAPL molecule.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / chemistry
  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / metabolism*
  • Amino Acid Sequence
  • Cell Membrane / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Lysosomes / metabolism
  • Protein Sorting Signals / genetics
  • Protein Structure, Secondary
  • Protein Structure, Tertiary / genetics

Substances

  • ABCB9 protein, human
  • ATP-Binding Cassette Transporters
  • Protein Sorting Signals
  • Green Fluorescent Proteins