2009
DOI: 10.1074/jbc.m109.044925
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Human Copper Transporter 1 Lacking O-Linked Glycosylation Is Proteolytically Cleaved in a Rab9-positive Endosomal Compartment

Abstract: The human copper transporter hCTR1 is a homotrimer composed of a plasma membrane protein of 190 amino acids that contains three transmembrane segments. The extracellular 65-amino acid amino terminus of hCTR1 contains both N-linked (at Asn 15 ) and O-linked (at Thr 27 ) sites of glycosylation. If O-glycosylation at Thr 27 is prevented, hCTR1 is efficiently cleaved, removing ϳ30 amino acids from the amino terminus. We have now investigated (i) the site of this cleavage, determining which peptide bonds are cleave… Show more

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Cited by 28 publications
(38 citation statements)
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“…5). Immunoblotting with an antibody raised against the Ctr1 amino terminus (30) showed cross-reactivity with the 35-37-kDa glycosylated form of Ctr1 and not the ϳ17-kDa amino-terminally truncated form (Fig. 5D), supporting the assignment of these polypeptide species.…”
Section: Posttranslational Control Of Ctr1 In Response Tomentioning
confidence: 64%
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“…5). Immunoblotting with an antibody raised against the Ctr1 amino terminus (30) showed cross-reactivity with the 35-37-kDa glycosylated form of Ctr1 and not the ϳ17-kDa amino-terminally truncated form (Fig. 5D), supporting the assignment of these polypeptide species.…”
Section: Posttranslational Control Of Ctr1 In Response Tomentioning
confidence: 64%
“…5C). Although exposure of HEK293T cells to copper resulted in clear reductions in the steady-state levels of the full-length glycosylated form of Ctr1, steady-state levels of a previously characterized Ctr1 species that was truncated 29 -33 residues from the amino terminus (25,30) were not significantly altered in response to any metal tested (Fig. 5).…”
Section: Posttranslational Control Of Ctr1 In Response Tomentioning
confidence: 98%
“…Confocal Imaging-Cells were grown on glass coverslips and treated in 12-well culture plates prior to fixing and staining with antibodies, exactly as described previously (29). Images of HEK293 cells were collected using a Zeiss LSM 700 confocal microscopy system equipped with a 100ϫ objective, controlled with Zen TM software.…”
Section: Methodsmentioning
confidence: 99%
“…The H139R and C-terminal mutants were derived from the FLAGtagged hCTR1 clone pEM94 (Ref. 29, available on request). H139R was made as described previously (29) with a site-directed mutagenesis kit (Stratagene, Carlsbad CA).…”
Section: Methodsmentioning
confidence: 99%
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