ABCC1 p.Ile301Ala
| Predicted by SNAP2: | A: N (82%), C: N (82%), D: N (78%), E: N (93%), F: N (87%), G: N (82%), H: N (82%), K: N (93%), L: N (97%), M: N (97%), N: N (87%), P: N (82%), Q: N (93%), R: N (93%), S: N (93%), T: N (93%), V: N (87%), W: N (57%), Y: N (87%), |
| Predicted by PROVEAN: | A: N, C: N, D: D, E: D, F: N, G: D, H: D, K: D, L: N, M: N, N: D, P: D, Q: D, R: D, S: N, T: N, V: N, W: D, Y: N, |
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[hide] Involvement of a di-leucine motif in targeting of ... Biochem Biophys Res Commun. 2013 Nov 8;441(1):89-95. doi: 10.1016/j.bbrc.2013.10.013. Epub 2013 Oct 12. Emi Y, Harada Y, Sakaguchi M
Involvement of a di-leucine motif in targeting of ABCC1 to the basolateral plasma membrane of polarized epithelial cells.
Biochem Biophys Res Commun. 2013 Nov 8;441(1):89-95. doi: 10.1016/j.bbrc.2013.10.013. Epub 2013 Oct 12., [PMID:24129190]
Abstract [show]
Localization of ATP-binding cassette transporter isoform C1 (ABCC1) to the basolateral membrane of polarized cells is crucial for export of a variety of cellular metabolites; however, the mechanism regulating basolateral targeting of the transporter is poorly understood. Here we describe identification of a basolateral targeting signal in the first cytoplasmic loop domain (CLD1) of human ABCC1. Comparison of the CLD1 amino acid sequences from ABCC1 to ABCC2 revealed that ABCC1 possesses a characteristic sequence, E(295)EVEALI(301), which is comprised of a cluster of acidic glutamate residues followed by a di-leucine motif. This characteristic sequence is highly conserved among vertebrate ABCC1 orthologs and is positioned at a site that is structurally equivalent to the apical targeting signal previously described in ABCC2. Alanine scanning mutagenesis of this sequence in full-length human ABCC1 showed that both L(300) and I(301) residues were required for basolateral targeting of ABCC1 in polarized HepG2 and MDCK cells. Conversely, E(295), E(296), and E(298) residues were not required for basolateral localization of the transporter. Therefore, a di-leucine motif within the CLD1 is a basolateral targeting determinant of ABCC1.
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No. Sentence Comment
92 Two ABCC1-HA point mutants, L300A and I301A, displayed a mixed distribution pattern, indicating diminished basolateral targeting specificity.
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ABCC1 p.Ile301Ala 24129190:92:38
status: NEW93 In addition to their usual localization to the basolateral membrane, significant proportions of the L300A and I301A mutants were found in the apical vacuoles of every polarized HepG2 cell.
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ABCC1 p.Ile301Ala 24129190:93:110
status: NEW94 All other ABCC1 point mutants tested exhibited normal basolateral localization, which excludes the possibility that perturbed localization of the L300A and I301A mutants was caused by gross protein misfolding arising from a point mutation and inhibition of ABCC1 trafficking to the basolateral membrane.
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ABCC1 p.Ile301Ala 24129190:94:156
status: NEW103 Similarly, MCCs of the I301A mutant were 0.63 &#b1; 0.05 and 0.59 &#b1; 0.04, respectively, which also suggests a mixed localization of the I301A mutant (Fig. 3C).
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ABCC1 p.Ile301Ala 24129190:103:23
status: NEWX
ABCC1 p.Ile301Ala 24129190:103:140
status: NEW114 Only two variants, L300A and I301A, exhibited a mixed localization pattern.
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ABCC1 p.Ile301Ala 24129190:114:29
status: NEW120 The L300A and I301A mutants lost their basolateral-specific targeting and produced a mixed staining pattern at both the apical and basolateral sides (Fig. 4).
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ABCC1 p.Ile301Ala 24129190:120:14
status: NEW129 HepG2 cells were transiently cotransfected with 0.5 lg of the wild-type (A), L300A (B), or I301A (C) ABCC1-HA expression plasmids, along with 0.5 lg of empty vector pCMV-HA.
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ABCC1 p.Ile301Ala 24129190:129:91
status: NEW151 Single merged panels of representative apical, lateral, and x-z sections are shown. The L300A and I301A mutants displayed a mixed localization to the apical and basolateral membranes.
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ABCC1 p.Ile301Ala 24129190:151:98
status: NEW113 Representative merged immunofluorescence images of the transfected cells are shown. The positions of apical vacuoles formed between juxtaposed polarized cells are indicated by white arrowheads. Scale bar, 20 lm. Only two variants, L300A and I301A, exhibited a mixed localization pattern.
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ABCC1 p.Ile301Ala 24129190:113:241
status: NEW119 The L300A and I301A mutants lost their basolateral-specific targeting and produced a mixed staining pattern at both the apical and basolateral sides (Fig. 4).
X
ABCC1 p.Ile301Ala 24129190:119:14
status: NEW128 HepG2 cells were transiently cotransfected with 0.5 lg of the wild-type (A), L300A (B), or I301A (C) ABCC1-HA expression plasmids, along with 0.5 lg of empty vector pCMV-HA.
X
ABCC1 p.Ile301Ala 24129190:128:91
status: NEW150 Single merged panels of representative apical, lateral, and x-z sections are shown. The L300A and I301A mutants displayed a mixed localization to the apical and basolateral membranes.
X
ABCC1 p.Ile301Ala 24129190:150:98
status: NEW