ABCMdb

PMID: 12034727

Mao Q, Qiu W, Weigl KE, Lander PA, Tabas LB, Shepard RL, Dantzig AH, Deeley RG, Cole SP
GSH-dependent photolabeling of multidrug resistance protein MRP1 (ABCC1) by [125I]LY475776. Evidence of a major binding site in the COOH-proximal membrane spanning domain.
J Biol Chem. 2002 Aug 9;277(32):28690-9. Epub 2002 May 28., 2002-08-09 [PubMed]

Sentences

No. Mutations Sentence Comment

56 ABCC1 p.Trp1246Ala ABCC1 p.Trp1246Ala ABCC1 p.Trp1246Phe ABCC1 p.Trp1246Phe ABCC1 p.Trp1246Tyr ABCC1 p.Trp1246Tyr Cell Culture and Membrane Protein Preparation-The doxorubicin-selected, multidrug-resistant H69AR small cell lung cancer cell line that expresses high levels of MRP1, and the transfected HeLa cell lines that express recombinant wild-type MRP1 (T5 or WT-MRP1), and mutant MRP1 bearing substitutions of Trp1246 (W1246F-MRP1, W1246Y-MRP1, W1245C-MRP1, and W1246A-MRP1) were maintained as described previously (15, 37). Login to comment 154 ABCC1 p.Trp1246Cys ABCC1 p.Trp1246Ala ABCC1 p.Trp1246Phe ABCC1 p.Trp1246Tyr After normalizing expression levels of the mutant MRP1 molecules relative to wild-type MRP1 (Fig. 8B), it was estimated that [125 I]LY475776 labeling of HeLa cell membrane proteins containing the W1246F-MRP1 mutant was ϳ23% of wild-type MRP1 membrane proteins, whereas labeling of membranes containing the W1246Y-MRP1, W1246C-MRP1, and W1246A-MRP1 mutants was less than 10% of wild-type MRP1 levels. Login to comment 163 ABCC1 p.Trp1246Cys ABCC1 p.Trp1246Ala ABCC1 p.Trp1246Phe ABCC1 p.Trp1246Tyr Membrane vesicles (50 ␮g of protein) were prepared from stably transfected HeLa cells expressing wild-type (WT-MRP1) and mutant MRP1 molecules in which Trp1246 has been replaced with Ala (W1246A), Phe (W1246F), Cys (W1246C), or Tyr (W1246Y). Login to comment