ABCMdb

PMID: 24833118

Engel T, Fobker M, Buchmann J, Kannenberg F, Rust S, Nofer JR, Schurmann A, Seedorf U
3beta,5alpha,6beta-Cholestanetriol and 25-hydroxycholesterol accumulate in ATP-binding cassette transporter G1 (ABCG1)-deficiency.
Atherosclerosis. 2014 Jul;235(1):122-9. doi: 10.1016/j.atherosclerosis.2014.04.023. Epub 2014 May 1., [PubMed]

Sentences

No. Mutations Sentence Comment

37 ABCG1 p.Lys124Met Cells TetOff-HeLa cell lines TG1-94 and TG1KM-47, stably transfected with pTRE expression plasmids (Clontech) containing wild-type (wt) or K124M-mutant (HA)3-ABCG1(666), respectively [13], were maintained in complete medium (Dulbecco`s modification of Eagle`s medium (DMEM) with 10% fetal calf serum (FCS), antibiotic-antimycotic solution, and glutamine), as previously described [13,14]. Login to comment 106 ABCG1 p.Lys124Met Analysis of ATPase activation by various oxysterols To address the question whether oxysterol species accumulating in Abcg1 deficiency are direct substrates for this transporter, individual oxysterols were applied to tetracycline-inducible HeLa cell lines expressing wt ABCG1 or mutant ABCG1-K124M variant, which was previously shown to be metabolically inactive [13,25]. Login to comment 112 ABCG1 p.Lys124Met As shown in Fig. 2, 25-HC (Fig. 2B, upper right panel), CT (Fig. 2B, lower left panel), and 7b-HC (Fig. 2B, lower right panel) induced MMP in wt ABCG1-expressing cells only (Fig. 2B bold lines), whereas no such effect could be noted in the cells expressing mutant ABCG1-K124M variant (Fig. 2B dashed lines). Login to comment 123 ABCG1 p.Lys124Met As shown in Fig. 4A, export of 25-HC was enhanced significantly in wt ABCG1-expressing cells, whereas cells expressing mutant ABCG1-K124M were ineffective in this respect. Login to comment 163 ABCG1 p.Lys124Met Expression of exogenous wt and mutant (K124M) HA3-ABCG1, as well as endogenous ABCG1 in a membrane fraction of cells grown as in (Fig. 4A and B) is shown in (A). Login to comment 167 ABCG1 p.Lys124Met (B) TetOff HeLa cell lines expressing (bold/dashed line) or not expressing (regular/dotted line) wt ABCG1 (bold/regular line) or mutant ABCG1-K124M (dashed/dotted line), were incubated in the presence of vehicle only (top left panel) or either 5 mM 25-HC (top right panel), 14.2 mM CT (bottom left panel), or 10 mM 7b-HC (bottom right panel), for 24 h. TMRE fluorescence was recorded (see Material and Methods). Login to comment