ABCMdb

PMID: 11741902

Hou YX, Cui L, Riordan JR, Chang XB
ATP binding to the first nucleotide-binding domain of multidrug resistance protein MRP1 increases binding and hydrolysis of ATP and trapping of ADP at the second domain.
J Biol Chem. 2002 Feb 15;277(7):5110-9. Epub 2001 Dec 7., 2002-02-15 [PubMed]

Sentences

No. Mutations Sentence Comment

50 ABCC1 p.Asp1454Leu ABCC1 p.Lys684Leu ABCC1 p.Glu1455Leu ABCC1 p.Lys1333Leu ABCC1 p.Asp792Ala Stable cell lines expressing wild-type and mutant MRP1s, K684L, D792A, K1333L, and D1454L/E1455L were established previously (2, 31). Login to comment 117 ABCC1 p.Asp1454Leu ABCC1 p.Lys684Leu ABCC1 p.Glu1455Leu ABCC1 p.Lys1333Leu ABCC1 p.Asp792Ala Essentially the stimulation was much reduced in the NBD1 mutants, K684L and D792A (Fig. 4, C and D), and in the NBD2 mutants, K1333L and D1454L/E1455L (Fig. 4, E and F), and the stimulation effects were shifted to higher ATP concentrations (Fig. 4, C-F). Login to comment 118 ABCC1 p.Asp1454Leu ABCC1 p.Asp1454Leu ABCC1 p.Glu1455Leu ABCC1 p.Glu1455Leu ABCC1 p.Lys1333Leu ABCC1 p.Lys1333Leu Trypsin digestion of either [␣-32 P]8-N3ATP or [␣-32 P]8N3ADP-labeled K1333L and D1454L/E1455L proved that the mutated NBD2 fragment can still be labeled (data not shown). Login to comment 173 ABCC1 p.Asp1454Leu ABCC1 p.Lys684Leu ABCC1 p.Glu1455Leu ABCC1 p.Lys1333Leu ABCC1 p.Asp792Ala Lane 1, 10 ␮g of wild-type MRP1; lane 2, 15 ␮g of K684L; lane 3, 20 ␮g of D792A; lane 4, 10 ␮g of K1333L; lane 5, 10 ␮g of D1454L/E1455L. Login to comment 175 ABCC1 p.Asp1454Leu ABCC1 p.Lys684Leu ABCC1 p.Lys684Leu ABCC1 p.Glu1455Leu ABCC1 p.Lys1333Leu ABCC1 p.Asp792Ala The results for K684L and D792A are the average of three independent experiments and for K1333L and D1454L/E1455L are the average of two independent experiments. C, influence of ATP on the [␣-32 P]8-N3ADP labeling of K684L. Login to comment 176 ABCC1 p.Lys684Leu ABCC1 p.Asp792Ala 15 ␮g of K684L was labeled in the presence of varying amounts of ATP indicated above each lane. D, influence of ATP on the [␣-32 P]8-N3ADP labeling of D792A. Login to comment 177 ABCC1 p.Asp792Ala 20 ␮g of D792A was labeled in the presence of varying amount of ATP indicated above each lane. Login to comment 178 ABCC1 p.Lys1333Leu E, influence of ATP on the [␣-32 P]8-N3ADP labeling of K1333L. Login to comment 179 ABCC1 p.Lys1333Leu 10 ␮g of K1333L was labeled in the presence of varying amounts of ATP indicated above each lane. Login to comment 180 ABCC1 p.Asp1454Leu ABCC1 p.Glu1455Leu F, influence of ATP on the [␣-32 P]8-N3ADP labeling of D1454L/E1455L. Login to comment 181 ABCC1 p.Asp1454Leu ABCC1 p.Glu1455Leu 10 ␮g of D1454L/E1455L was labeled in the presence of varying amounts of ATP indicated above each lane. Login to comment