ABCMdb

ABCC1 p.Pro104Ala

Predicted by SNAP2:	A: N (72%), C: N (72%), D: N (57%), E: D (59%), F: D (53%), G: N (53%), H: N (72%), I: N (53%), K: D (63%), L: N (57%), M: D (53%), N: N (61%), Q: N (66%), R: D (59%), S: N (72%), T: N (66%), V: N (57%), W: N (53%), Y: N (61%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: D, K: D, L: D, M: D, N: D, Q: D, R: D, S: D, T: D, V: D, W: D, Y: D,

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Publications
[hide] Mutation of proline residues in the NH(2)-terminal... Biochim Biophys Acta. 2003 Sep 2;1615(1-2):103-14. Ito K, Weigl KE, Deeley RG, Cole SP
Mutation of proline residues in the NH(2)-terminal region of the multidrug resistance protein, MRP1 (ABCC1): effects on protein expression, membrane localization, and transport function.
Biochim Biophys Acta. 2003 Sep 2;1615(1-2):103-14., [PMID:12948592]

Abstract [show]
The Multidrug Resistance Protein, MRP1 (ABCC1) confers drug resistance and transports organic anions such as leukotriene C(4) (LTC(4)) and 17beta-estradiol 17-(beta-D-glucuronide) (E(2)17betaG). Previous studies showed that portions of the first membrane spanning domain (MSD1) and the cytoplasmic loop (CL3) connecting it to MSD2 are important for MRP1 transport function. We have replaced 12 prolines in MSD1 and CL3 with alanine and determined the effects of these substitutions on MRP1 expression and transport activity. All singly substituted MRP1-Pro mutants could be expressed in HeLa cells, except MRP1-P104A. The expressed mutants also transported LTC(4) and E(2)17betaG, and their K(m) (LTC(4)) values were similar to wild-type MRP1. Expression of the double mutant MRP1-P42/51A was reduced by >80% although it localized to the plasma membrane and transported organic anions. MRP1 expression was also reduced when the first transmembrane helix (amino acids 37-54) was deleted. In contrast, the phenotypes of the multiply substituted CL3 mutants MRP1-P196/205/207/209A and MRP1-P235/255A were comparable to wild-type MRP1. However, Pro(255)-substituted MRP1 mutants showed reduced immunoreactivity with a monoclonal antibody (MAb) whose epitope is located in CL3. We conclude that certain prolines in MSD1 and CL3 play a role in the expression and structure of MRP1.

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No. Sentence Comment
3 All singly substituted MRP1-Pro mutants could be expressed in HeLa cells, except MRP1-P104A. Login to comment
46 For the single substitutions of Pro with Ala in MSD1, the following sense mutagenic primers were used (substituted nucleotides are underlined): MRP1-P42A (5V-C GTG TGG GTG GCT TGT TTT TAC CTC TGG-3V), MRP1-P51A (5V-G GCC TGT TTC GCC TTC TAC TTC C-3V), MRP1-P69A (5V-CAG ATG ACA GCT CTC AAC-3V), MRP1-P104A (5V-C ATA TTC CTG GCC GCA GTG TTT CTG G-3V), MRP1-P110A (5V-GT TTC TGG TCA GCG CAA CTC TCT TGG-3V). Login to comment
103 Despite multiple attempts, stable HeLa cell lines expressing MRP1-P104A and MRP1-P104/110A could not be established. Login to comment