RGD Reference Report - Vesicular localization of the rat ATP-binding cassette half-transporter rAbcb6. - Rat Genome Database

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Vesicular localization of the rat ATP-binding cassette half-transporter rAbcb6.

Authors: Jalil, Youssef Abdul  Ritz, Vera  Jakimenko, Ana  Schmitz-Salue, Christoph  Siebert, Heike  Awuah, David  Kotthaus, André  Kietzmann, Thomas  Ziemann, Christina  Hirsch-Ernst, Karen I 
Citation: Jalil YA, etal., Am J Physiol Cell Physiol. 2008 Feb;294(2):C579-90. doi: 10.1152/ajpcell.00612.2006. Epub 2007 Dec 26.
RGD ID: 42722002
Pubmed: PMID:18160489   (View Abstract at PubMed)
DOI: DOI:10.1152/ajpcell.00612.2006   (Journal Full-text)

The clarification of subcellular localization represents an important basis toward characterization of ATP-binding cassette (ABC) transporters and resolution of their roles in cellular physiology. Rat Abcb6 (rAbcb6) is a membrane-situated half-transporter belonging to the ABC protein superfamily. To investigate rAbcb6 subcellular distribution, the human colon adenocarcinoma line LoVo, which we found to be devoid of endogenous human ABCB6 mRNA, was employed for heterologous expression of rAbcb6 bearing a COOH-terminal epitope tag (rAbcb6-V5). Following subcellular fractionation, rAbcb6-V5 was observed as an N-glycosylated protein in fractions enriched with lysosomal/endosomal membrane proteins. Indirect immunofluorescence analyses of rAbcb6-V5 using antibodies against a rAbcb6-specific peptide or against the V5-tag revealed a punctate pattern that was colocalized with lysosome-associated membrane protein 1 (LAMP1), a marker of lysosomes/late endosomes. Substantial colocalization of tagged rAbcb6 with lysosomal/late endosomal marker was confirmed with living, unfixed LoVo cells coexpressing rAbcb6 fused to enhanced green fluorescent protein. Vesicular distribution in LoVo cells was consistent with localization of endogenous rAbcb6 expressed in rat primary hepatocyte cultures or in liver sections, as revealed by overlap of rat Lamp1 with rAbcb6 in double immunofluorescence analyses. Since several Abcb6-related half-transporters confer heavy metal tolerance, we investigated whether rAbcb6 expression in LoVo cells might affect sensitivity toward transition metal toxicity. Applying MTT viability assays, we found that expression of either rAbcb6-V5 or untagged rAbcb6 conferred tolerance toward copper, but not to cobalt or zinc. In summary, these results demonstrate that rAbcb6 is a glycosylated protein targeted to intracellular vesicular membranes and suggest involvement of rAbcb6 in transition metal homeostasis.

Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
intracellular copper ion homeostasis involved_inIDA 42722002PMID:18160489UniProt 

Cellular Component
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
early endosome membrane located_inIDA 42722002PMID:18160489UniProt 
endolysosome membrane located_inIDA 42722002PMID:18160489UniProt 
late endosome membrane located_inIDA 42722002PMID:18160489UniProt 
lysosomal membrane located_inIDA 42722002PMID:18160489UniProt 

Objects Annotated

Genes (Rattus norvegicus)
Abcb6  (ATP binding cassette subfamily B member 6)


Additional Information