ABCMdb

ABCC1 p.Leu370Val

Predicted by SNAP2:	A: D (63%), C: D (53%), D: D (85%), E: D (80%), F: N (72%), G: D (80%), H: D (80%), I: N (78%), K: D (85%), M: N (93%), N: D (80%), P: D (85%), Q: D (75%), R: D (85%), S: D (71%), T: D (75%), V: D (53%), W: D (75%), Y: D (59%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: N, K: D, M: N, N: D, P: D, Q: D, R: D, S: D, T: D, V: N, W: D, Y: D,

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Publications
[hide] ATPase activity of purified multidrug resistance-a... J Biol Chem. 1997 Dec 5;272(49):30962-8. Chang XB, Hou YX, Riordan JR
ATPase activity of purified multidrug resistance-associated protein.
J Biol Chem. 1997 Dec 5;272(49):30962-8., [PMID:9388243]

Abstract [show]
Human multidrug resistance protein (MRP) was expressed at high levels in stably transfected baby hamster kidney (BHK-21) cells. These cells exhibited a pattern of cross-resistance to several different drugs typical of an MRP-mediated phenotype despite the addition of 10 histidine residues at the C terminus to facilitate purification. Consistent with this functional evidence of the presence of MRP at the surface of these transfectants, strong signals were detected by immunoblotting and immunofluorescence using a specific monoclonal antibody to MRP. There was intense uniform staining of the cell surface as well as weaker staining of intracellular membranes. MRP-containing membranes were solubilized in 1% N-dodecyl-beta-D-maltoside in the presence of 0.4% sheep brain phospholipids. Two sequential affinity purification steps on Ni-NTA agarose and wheat germ agglutinin agarose provided substantial enrichment, and contaminating bands were not detected. ATPase activity of the purified protein was assayed in the presence of the phospholipids, which had been maintained throughout all purification steps. ATP was hydrolyzed in proportion to the amount of purified protein assayed, and typical Michaelis-Menten behavior was exhibited, yielding estimations of Km of approximately 3.0 mM and Vmax of 0.46 micromol mg-1 min-1. This activity was moderately stimulated by the drugs that others have shown to be transported by MRP-containing membrane vesicles. This stimulation was enhanced by reduced glutathione as is its drug transport, and oxidized glutathione, itself a substrate for transport, caused a strong stimulation. These data describe the first purification of MRP and provide the first direct evidence that the molecule possesses drug-stimulated ATPase activity.

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No. Sentence Comment
93 These differences and the resulting amino acid changes are as follows: Thr434 to Gly (L80V), Cys546 to Thr (T117M), Cys1304 to Gly (L370V), Thr1880 to Cys (L582L), Thr2250 to Cys (L685S), Ala2283 to Gly (D696G), Cys4040 to Gly and Gly4041 to Cys (R1282A), and Gly4732 to Ala (S1512S). Login to comment
105 L370V represents a conservative hydrophobic replacement in TM2 of the currently favored topology model (31). Login to comment
91 These differences and the resulting amino acid changes are as follows: Thr434 to Gly (L80V), Cys546 to Thr (T117M), Cys1304 to Gly (L370V), Thr1880 to Cys (L582L), Thr2250 to Cys (L685S), Ala2283 to Gly (D696G), Cys4040 to Gly and Gly4041 to Cys (R1282A), and Gly4732 to Ala (S1512S). Login to comment
103 L370V represents a conservative hydrophobic replacement in TM2 of the currently favored topology model (31). Login to comment