ABCC1 p.Cys388Asp
| Predicted by SNAP2: | A: N (66%), D: D (80%), E: D (80%), F: D (75%), G: N (57%), H: D (80%), I: N (53%), K: D (85%), L: D (63%), M: N (66%), N: D (63%), P: D (85%), Q: D (80%), R: D (85%), S: N (61%), T: N (61%), V: N (53%), W: D (80%), Y: D (80%), |
| Predicted by PROVEAN: | A: D, D: D, E: D, F: D, G: D, H: D, I: D, K: D, L: D, M: D, N: D, P: D, Q: D, R: D, S: D, T: D, V: D, W: D, Y: D, |
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[hide] Effect of multiple cysteine substitutions on the f... Drug Metab Dispos. 2012 Jul;40(7):1403-13. Epub 2012 Apr 16. Qin L, Tam SP, Deeley RG
Effect of multiple cysteine substitutions on the functionality of human multidrug resistance protein 1 expressed in human embryonic kidney 293 cells: identification of residues essential for function.
Drug Metab Dispos. 2012 Jul;40(7):1403-13. Epub 2012 Apr 16., [PMID:22511347]
Abstract [show]
Multidrug resistance protein 1 (MRP1) is a broad-specificity membrane transporter belonging to the C branch of the ATP binding cassette (ABC) superfamily. MRP1 confers resistance to various chemotherapeutic drugs and transports a wide range of conjugated organic anions. Several ABCC proteins, including MRP1, are unusual among ABC transporters in having a third membrane-spanning domain (MSD), MSD0, at their N termini. MRP1 lacking this additional MSD (DeltaMRP1) is able to traffic to the plasma membrane of mammalian cells and to transport a number of well characterized substrates. A cysteineless (cysless) DeltaMRP1 has been expressed in yeast and reported to be functional. However, we found that trafficking of such a construct in human cells was severely compromised, and, even when expressed in insect Sf21 cells, the protein had extremely low transport activity. Therefore, we have systematically examined the effects of substituting cysteines in the four domains of DeltaMRP1, initially with alanine. These studies allowed us to identify five cysteines that cannot be replaced with alanine without inactivating the protein. Substitution of two of these residues with alternative amino acids has allowed us to produce an almost cysless form of DeltaMRP1 that traffics to the plasma membrane and transports leukotriene C(4), 17beta-estradiol 17-beta-D-glucuronide, and estrone-3-sulfate with kinetic characteristics similar to those of the wild-type protein. The distribution of the remaining Cys residues is such that the protein will provide a useful template for a variety of cysteine based mutagenesis studies.
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No. Sentence Comment
331 Substitution of Cys388 with seven different amino acids (C388D, C388G, C388I, C388M, C388N, C388S, and C388V) decreased the transport of LTC4 by 30% (C388V) to more than 90% (C388N).
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ABCC1 p.Cys388Asp 22511347:331:57
status: NEW332 Their effect on E217betaG transport was similar although the C388V mutation reduced transport by only 10 to 15% (data not shown).
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ABCC1 p.Cys388Asp 22511347:332:57
status: NEW