ABCMdb

ABCG5 p.Asn591Gln

None has been submitted yet.

Publications

PMID: 24584735 [PubMed] Suzuki S et al: "Inhibition of post-translational N-glycosylation by HRD1 that controls the fate of ABCG5/8 transporter."

No. Sentence Comment

50 Single substitution of glutamine for asparagine at position 584 (N584Q) and at position 591 (N591Q) in human ABCG5 reduced the apparent molecular mass to a similar extent between two mutants (Figure 1b). Login to comment
51 Double mutation (N584Q/N591Q) further reduced the band size identical to those of EndoH- and PNGase F-treated ABCG5 proteins (Figure 1b,c; Supplementary Figure S4), indicating that human ABCG5 contains two N-linked glycans at positions 584 and 591. Login to comment
67 (b) Steady-state expression of monomeric Myc-tagged WT-, N584Q-, N591Q- and N584Q-/N591Q-ABCG5 was analyzed by immunoblotting. Login to comment
68 (c) Endo H (500 U) and PNGase F (500 U) sensitivity of Myc-tagged WT-, N584Q-, N591Q- and N584Q-/N591Q-ABCG5 proteins shown in (b). Login to comment
83 (f-i) Pulse chase (pulse labeled for 10 min) of HEK293 cells expressing Myc-tagged WT-, N584Q- and N591Q-ABCG5 (f). Login to comment
84 Relative quantity of each glycosylated form of WT-ABCG5 (g), N591Q-ABCG5 (h) and N584Q-ABCG5 (i) within all ABCG5 was quantified. Login to comment
91 To further characterize the glycosylation timing of each Asn residues (N584 and N591) of ABCG5 protein, mutational analysis using N584Q- and N591Q-ABCG5 constructs were performed. Login to comment
92 Notably, Non-G forms of both N584Q and N591Q ABCG5 proteins were abundantly expressed (Figure 2f,h,i: 70.1% vs. 54.6%), indicating that both Asn residues (N584 and N591) have the potential to be post-translationally glycosylated although the tendency seems to be different among residues. Login to comment
93 Consistently, pulse-chase analysis using N584Q and N591Q ABCG5 revealed that the expression level of glycosylated forms of N584Q and N591Q ABCG5 are gradually increased during the chase period (Figure 2f,h,i: 45.4% to 72.6% for N584Q-ABCG5; 29.9% to 71.9% for N591Q-ABCG5). Login to comment
94 Because of the high expression of Non-G form of N591Q ABCG5 at 0-min chase (Figure 2f,h 70.1%) and of the time-dependent increase of Mono-G form of ABCG5 (Figure 2f,h), we can deduce that most of the N584 residue of ABCG5 is post-translationally glycosylated, while N591 residue is both co- and post-translationally glycosylated. Login to comment
302 Glycosylation-defective mutants N584Q-, N591Q-, N584Q/N591Q-ABCG5, N619Q-ABCG8, and E3-defective mutants C291S/C329S-HRD1, H260Q CHIP and C42S-RMA1 were generated using the QuikChange II site-directed mutagenesis kit (Stratagene, La Jolla, CA) according to previous reports12,17,32-34 . Login to comment