ABCC1 p.Tyr324Ala
| Predicted by SNAP2: | A: D (59%), C: D (53%), D: D (71%), E: D (63%), F: N (61%), G: D (71%), H: N (57%), I: N (57%), K: D (71%), L: N (53%), M: N (57%), N: D (66%), P: D (66%), Q: D (53%), R: D (63%), S: D (53%), T: D (53%), V: N (57%), W: N (53%), |
| Predicted by PROVEAN: | A: D, C: D, D: D, E: D, F: N, 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, |
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[hide] Molecular modeling of the human multidrug resistan... Biochem Biophys Res Commun. 2008 Jan 4;365(1):29-34. Epub 2007 Oct 31. DeGorter MK, Conseil G, Deeley RG, Campbell RL, Cole SP
Molecular modeling of the human multidrug resistance protein 1 (MRP1/ABCC1).
Biochem Biophys Res Commun. 2008 Jan 4;365(1):29-34. Epub 2007 Oct 31., 2008-01-04 [PMID:17980150]
Abstract [show]
Multidrug resistance protein 1 (MRP1/ABCC1) is a 190kDa member of the ATP-binding cassette (ABC) superfamily of transmembrane transporters that is clinically relevant for its ability to confer multidrug resistance by actively effluxing anticancer drugs. Knowledge of the atomic structure of MRP1 is needed to elucidate its transport mechanism, but only low resolution structural data are currently available. Consequently, comparative modeling has been used to generate models of human MRP1 based on the crystal structure of the ABC transporter Sav1866 from Staphylococcus aureus. In these Sav1866-based models, the arrangement of transmembrane helices differs strikingly from earlier models of MRP1 based on the structure of the bacterial lipid transporter MsbA, both with respect to packing of the twelve helices and their interactions with the nucleotide binding domains. The functional importance of Tyr324 in transmembrane helix 6 predicted to project into the substrate translocation pathway was investigated.
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No. Sentence Comment
34 Consequently, we have tested the functional importance of this residue by using site-directed mutagenesis to replace Tyr324 with Ala, Phe and Trp, and characterizing the transport activity of the resulting mutants.
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ABCC1 p.Tyr324Ala 17980150:34:117
status: NEW46 To create the single mutants Y324A, Y324F and Y324W, the QuikChangeä site-directed mutagenesis kit (Stratagene Inc., La Jolla, CA), was used.
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ABCC1 p.Tyr324Ala 17980150:46:29
status: NEW48 The mutations were introduced using the following sense primers (mutated residues are underlined, silent mutations to introduce restriction sites are italicized, and restriction enzymes used are indicated in parentheses): Y324A, 50 -GAAGAAGCT CATGAGGAATGCGGGCCCAAAGGTC-30 (BsmI); Y324F, 50 -GCT CATGAGGAAGAACGGCCCAAAGGTCTTG-30 (ApaI); Y324W, 50 - GAAGAAGCTCATCAGGAACCAGGGCCCAAAGGTC-30 (BspHI).
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ABCC1 p.Tyr324Ala 17980150:48:222
status: NEW