ABCMdb

PMID: 16870176

Engel T, Bode G, Lueken A, Knop M, Kannenberg F, Nofer JR, Assmann G, Seedorf U
Expression and functional characterization of ABCG1 splice variant ABCG1(666).
FEBS Lett. 2006 Aug 7;580(18):4551-9. Epub 2006 Jul 13., [PubMed]

Sentences

No. Mutations Sentence Comment

138 ABCG1 p.Lys124Met Both, lipoprotein particles as well as serum albumin take up cholesterol twice as efficiently from wt ABCG1 expressing cells as compared to the same cell line where ABCG1 expression had been turned off by application of doxycycline, the parental TetOff HeLa cells or a cell line expressing inactive mutant ABCG1(666; K124M). Login to comment 170 ABCG1 p.Lys124Met ABCG1(666; K124M) mutant localizes at the cell surface and expression can be regulated by doxycycline. Login to comment 171 ABCG1 p.Lys124Met TetOff HeLa cells were stable transfected with plasmids coding for wt (HA)3-ABCG1(666), mutant (HA)3-ABCG1(666; K124M) or (HA)3-ABCG2, respectively. Login to comment 174 ABCG1 p.Lys124Met (B) Lanes 5 and 6: (HA)3-ABCG1(666; K124M) clone 47. Login to comment 184 ABCG1 p.Lys124Met [3 H]cholesterol efflux to various acceptors was measured for 1.5 h in TetOff HeLa cells either stably transfected with (HA)3-ABCG1(666) (G1-94; white, black), (HA)3-ABCG1(666; K124M) (G1(KM)-47; light/dark grey), or non-transfected (TetOff-HeLa; cross-hatched) in the presence (white, light gray, hatched) or absence (black, dark gray, hatched gray) of 1 lg/ml doxycycline, as described in Section 2. differed from previously characterized ABCG1 variants by absence of an internal segment of 12 amino acids (Fig. 8). Login to comment 204 ABCG1 p.Lys124Met (A) Samples from TetOff HeLa cell lines expressing wt or mutant (K124M) (HA)3- ABCG transporter, as indicated, were subjected to reducing (right panel) or non-reducing (left panel) SDS-PAGE and subsequent Western blot analysis with HA.11 antibody followed by goat anti-mouse HRP-labeled secondary antibody. Login to comment 206 ABCG1 p.Lys124Met G1-94, (HA)3-ABCG1(666) clone 94; G1KM-47, (HA)3-ABCG1(666, K124M) clone 47; G2-63, (HA)3-ABCG2 clone 63. Login to comment 235 ABCG1 p.Lys124Met We thank Dr. Helen Hobbs (UTSW, Dallas, TX, USA) for providing us with an ABCG5-(FLAG)3 plasmid used as a template for cloning ABCG5 employed in control 1_HSG1A 1 SDHKRDLGGDAEVNPFLWHRPSEE------------DSSSMEGCHSFSASCLTQFCILFKRTFL 2_HSG1B 1 SDHKRDLGGDAEVNPFLWHRPSEEVKQTKRLKGLRKDSSSMEGCHSFSASCLTQFCILFKRTFL 3_MMG1_ 1 ADYKRDLGGDTDVNPFLWHRPAEE------------DSASMEGCHSFSASCLTQFCILFKRTFL 4_RNG1_ 1 SDYKRELGGDGDVNPFLWHRPAEE------------DSASMEGCHSFSASCLTQFCILFKRTFL 6_CFG1_ 1 SEHRREPGGDAEVNPFLWHRPSEE------------DSTSMEGCHSFSASCLTQFCILFKRTFL 5_BTG1_ 1 SDCRREPGGDAEVNPFIWHRPSEE------------DSTSMEGCHSFSASCLTQFCILFKRTFL 7_DRG1_ 1 KDYKTEMNGNGVHNPFLWHRPSDE------------DSSSSEGCHSFSASCLTQFCILFKRTFL 8_HSG4_ 1 MAEKKSSPEKNEVPAPCPPCPPEV------------DPIES---HTFATSTLTQFCILFKRTFL hotloop hotloop ordered dis- HSG1A HSG1B A B COOH NH2 Walker B Signature, motif C Walker A outside cytoplasma K124M potential NGS * # 379-VKQTKRLKGLRK-390 Fig. 8. Login to comment 243 ABCG1 p.Lys124Met The mutated site in the Walker-A motif of the transporter, used for generating ABCG1(666; K124M) mutant, is indicated as well (#). Login to comment