ABCG2 p.Asn418Gln
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PMID: 15807535
[PubMed]
Diop NK et al: "N-Linked glycosylation of the human ABC transporter ABCG2 on asparagine 596 is not essential for expression, transport activity, or trafficking to the plasma membrane."
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32
In this study, we identified Asn 596 as the single N-linked glycosylated amino acid in ABCG2 using chemical treatments with tunicamycin or PNGase F and site-directed mutagenesis, constructing and evaluating three N f Q variants (N418Q, N557Q, and N596Q).
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ABCG2 p.Asn418Gln 15807535:32:229
status: VERIFIED52 The following genetic variants were obtained: Asn 418 (N418Q), Asn 557 (N557Q), and Asn 596 (N596Q).
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ABCG2 p.Asn418Gln 15807535:52:55
status: VERIFIED59 A 70-80% confluent monolayer of HeLa cells was infected with the vTF 7-3 vaccinia virus (10 pfu/cell) and cotransfected with an appropriate amount of a pTM1 expression plasmid containing ABCG2 or the following ABCG2 variants: N418Q, N557Q, and N596Q.
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ABCG2 p.Asn418Gln 15807535:59:226
status: VERIFIED153 Site-directed mutagenesis was performed on the human ABCG2 cDNA to generate three mutant variants of the half-transporter where the Asn residues were replaced by Gln residues: ABCG2 (N418Q), ABCG2 (N557Q), and ABCG2 (N596Q).
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ABCG2 p.Asn418Gln 15807535:153:183
status: VERIFIED156 ABCG2 (N418Q) and ABCG2 (N557Q) migrated as a range of bands between the 50 and 75 kDa markers and are indistinguishable from the glycosylated ABCG2 control (Figure 4A, lanes 1, 3, and 5).
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ABCG2 p.Asn418Gln 15807535:156:7
status: VERIFIED161 Upon treatment with PNGase F, both ABCG2 variants N418Q (Figure 4A, lane 4) and N557Q (Figure 4A, lane 6) also migrate to the same lower molecular mass position (~60 kDa) as the PNGase F treated ABCG2 glycosylated protein and the untreated N596Q protein (Figure 4A, lanes 2, 4, 6, and 7).
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ABCG2 p.Asn418Gln 15807535:161:50
status: VERIFIED181 The negative control membranes, containing the empty vector pTM1, show a basal activity of approximately 6.5 ( 1.6 nmol π min-1 mg-1 and FIGURE 4: Immunoblot detection of ABCG2 and the N418Q, N557Q, and N596Q ABCG2 variants.
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ABCG2 p.Asn418Gln 15807535:181:191
status: VERIFIED