ABCMdb

PMID: 25315773

Takatsu H, Tanaka G, Segawa K, Suzuki J, Nagata S, Nakayama K, Shin HW
Phospholipid flippase activities and substrate specificities of human type IV P-type ATPases localized to the plasma membrane.
J Biol Chem. 2014 Nov 28;289(48):33543-56. doi: 10.1074/jbc.M114.593012. Epub 2014 Oct 14., [PubMed]

Sentences

No. Mutations Sentence Comment

207 ABCB4 p.Lys435Met To address this hypothesis, we established cell lines stably expressing FLAG-tagged ABCB4(WT) or ABCB4(K435M), which is nonfunctional due to a mutation in the Walker A motif in the nucleotide-binding domain (41), with or without ATP8B1-HA. Login to comment 208 ABCB4 p.Lys435Met In both the singly and doubly expressing cell lines, comparable expression levels of ABCB4(WT) and ABCB4(K435M) were confirmed by immunoblotting, and the plasma membrane localization of all proteins was visualized by immunostaining with anti-FLAG antibody (Fig. 9, A and B). Login to comment 210 ABCB4 p.Lys435Met ABCB4(WT) and ABCB4- (K435M) colocalized with ATP8B1 at the plasma membrane (Fig. 8B). Login to comment 211 ABCB4 p.Lys435Met Importantly, coexpression of ATP8B1 with ABCB4- (WT), but not ABCB4(K435M), significantly suppressed PC incorporation at all time points examined relative to that in cells expressing only ATP8B1 (Fig. 9, C and D). Login to comment 214 ABCB4 p.Lys435Met Although we did not detect a decrease in PC incorporation in cells expressing ABCB4 alone at 5 min, we did detect a small but significant decrease at 30 min relative to parental cells or cells expressing ABCB4(K435M) (Fig. 9C). Login to comment 219 ABCB4 p.Lys435Met ABCB4 p.Lys435Met HeLa cell lines stably expressing HA-tagged ATP8B1, FLAG-tagged ABCB4(WT), FLAG-tagged ABCB4(K435M), ATP8B1-HA af9; FLAG-ABCB4(WT), or ATP8B1-HA af9; FLAG-ABCB4(K435M) were established by infection with recombinant retroviral vectors. Login to comment