ABCMdb

PMID: 11952788

Nies AT, Konig J, Cui Y, Brom M, Spring H, Keppler D
Structural requirements for the apical sorting of human multidrug resistance protein 2 (ABCC2).
Eur J Biochem. 2002 Apr;269(7):1866-76., [PubMed]

Sentences

No. Mutations Sentence Comment

138 ABCC2 p.Thr1543Ala We therefore deleted the C-terminal three amino acids or substituted threonine with alanine at position 1543. Login to comment 158 ABCC2 p.Thr1543Ala Asacontrol,localizationofGFP-MRP2,GFP-MRP2D3, and GFP-MRP2-T1543A was also analyzed in MDCKII cells grown polarized on Transwell filter membranes (Fig. 6). Login to comment 160 ABCC2 p.Thr1543Ala GFP-MRP2, GFP-MRP2D3, and GFP-MRP2-T1543A were almost exclusively present in the apical membrane with some GFP fluorescence also present in intracellular compartments. Login to comment 192 ABCC2 p.Thr1543Ala Because HepG2 cells endogenously Table 1. Quantitative analysis of the subcellular localization of C-terminally mutated GFP-MRP2 constructs in polarized HepG2 cells. Data are percentages of cells in which the respective localization of recombinant protein was observed as described in Materials and methods. Cells were observed 2 days after transfection. Data are means ±SD of six transient transfections using butyrate-induced cells as described under Materials and methods. Construct % Apical % Vesicles % ER C-Terminal sequence (1516-1545) GFP-MRP2 73 ± 9 18 ± 9 9 ± 5 GSPEELLQIPGPFYFMAKEAGIENVNSTKF GFP-MRP2D3 64 ± 9 13 ± 5 23 ± 9 GSPEELLQIPGPFYFMAKEAGIENVNS GFP-MRP2-T1543A 67 ± 6 16 ± 2 17 ± 6 GSPEELLQIPGPFYFMAKEAGIENVNSAKF GFP-MRP2D15 16 ± 7 17 ± 7 67 ± 14 GSPEELLQIPGPFYF GFP-MRP2D15TKF 21 ± 11 21 ± 7 58 ± 11 GSPEELLQIPGPFYFTKF Fig. 6. Login to comment 193 ABCC2 p.Thr1543Ala Localization of GFP-MRP2 (green in A,B), GFP-MRP2D3 (green in C,D), and GFP-MRP2-T1543A (green in E,F) in polarized MDCKII cells. Login to comment 197 ABCC2 p.Thr1543Ala The intense yellow color in the x-z planes, due to merging of the green GFP and the red concanavalin A fluorescence, shows that GFP-MRP2, GFP-MRP2D3, and GFP-MRP2-T1543A are almost exclusively localized in the apical membrane. Login to comment