ABCMdb

PMID: 9753453

Hrycyna CA, Airan LE, Germann UA, Ambudkar SV, Pastan I, Gottesman MM
Structural flexibility of the linker region of human P-glycoprotein permits ATP hydrolysis and drug transport.
Biochemistry. 1998 Sep 29;37(39):13660-73., 1998-09-29 [PubMed]

Sentences

No. Mutations Sentence Comment

83 ABCB1 p.Gly185Val pSX-MDR1/A is a pGEM3-based vector in which the multiple cloning site has been eliminated and replaced with a SacII-XhoI site to introduce a cDNA encoding the G185V MDR1 variant. Login to comment 111 ABCB1 p.Gly185Val ABCB1 p.Gly185Val Transfections followed by colchicine selection with the modified pSK4ADA-MDR1 (G185V) DNA (flexible linker experiments) and modified pHaMDR1/A (G185V) DNA (R-helical experiments) were performed similarly but using 100 mm dishes throughout. Login to comment 188 ABCB1 p.Gly185Val Table 1 demonstrates that similar numbers of drug-resistant colonies were obtained with unmodified MDR1 and MDR1-(flexible insertion), indicating that the modified transporter was approximately as effective as the G185V variant Pgp in conferring resistance to colchicine. Login to comment 191 ABCB1 p.Gly185Val The experiments described in Table 1 were performed with the G185V variant of Pgp, a well-characterized mutant that has been shown to change the substrate specificity of Pgp compared to that of the wild type, increasing its ability to confer resistance to colchicine (23). Login to comment 192 ABCB1 p.Gly185Val We found that the insertion mutant in Table 1 does not have an altered phenotype compared to the G185V variant, suggesting that the insertion of these additional amino acid residues did not affect the ability of the transporter to confer resistance to colchicine. Login to comment 200 ABCB1 p.Gly185Val To further examine the failure of the R-helical peptide insertion to confer drug resistance, parental G185V Pgp and the R-helical peptide insertion construct were transfected into NIH3T3 cells and the resultant transiently expressing populations were analyzed by FACS. Login to comment