ABCMdb

ABCG2 p.Cys374Ala

None has been submitted yet.

Publications

PMID: 18430864 [PubMed] Liu Y et al: "Effect of cysteine mutagenesis on the function and disulfide bond formation of human ABCG2."

No. Sentence Comment

108 To map which of the three cysteine residues are functionally important, we engineered four more constructs by mutating all or partial of the three cysteines Cys284, Cys374, and Cys438 to alanines and generated constructs I3-CL, I2-CL, C284A, and C374A (see Fig. 5A). Login to comment
111 Mutation of one or two of these residues (I2-CL, C284A, and C374A) did not signifi- TABLE 1 Primers used for construction of cysless mutants Mutations Primer Sequence RESa C43A TTTCATAACATTGCCTATCGAGTAAAACTGAAG BsrDI C55A GCTTTCTACCTGCACGAAAACCAGTTGAG BsgI C119A GCCAATTTCAAAGCGAATTCAGGTTACGTGG EcoRI C284A GAATCAGCTGGATATCACGCTGAGGCCTATAATAAC EcoRV C374A ACACCACCTCCTTCGCTCATCAACTCAGATG None C438A CTGACGACCAACCAAGCTTTCAGCAGTGTTTC HindIII C491A TATATTTACCGCTATAGTATACTTCATGTTAGG AccI C544A CTTCTCATGACGATCGCTTTTGTGTTTATGATG PvuI C592A GGACAAAACTTCGCCCCGGGACTCAATGCAA SmaI C603A/C608A AGGAAACAATCCTGCTAACTATGCAACAGCTACTGGCGAAGAATATTT -NspI C635A CACGTGGCCTTGGCTGCAATGATTGTTATTTTC BsrDI a Restriction (RES) enzyme digestion sites engineered in the primer for the convenience of detection. Login to comment
114 I3-CL with C284A, C374A, and C438A mutations also lost most of its ability to transport another substrate, Hoechst 33342 (Fig. 5D). Login to comment
150 C, relative activity of mutant ABCG2R482G I2-CL, C284A, I3-CL, and C374A compared with wild-type and ABCG2R482G as determined using flow cytometry for mitoxantrone (MX) accumulation in Sf9 cells. Data shown are from three independent experiments with S.D. D, effect of C284A, C374A, and C438A mutations (construct I3-CL) on efflux of Hoechst 33342 (Hoechst) as determined using flow cytometry. Login to comment
169 The observation that the single mutation of these two cysteine residues (C284A and C374A) did not substantially affect ABCG2R482G activity confirms the fact that their mutations did not severely influence the nucleotide-binding activity. Login to comment