ABCMdb

PMID: 15499164

Ishikawa T, Hirano H, Onishi Y, Sakurai A, Tarui S
Functional evaluation of ABCB1 (P-glycoprotein) polymorphisms: high-speed screening and structure-activity relationship analyses.
Drug Metab Pharmacokinet. 2004 Feb;19(1):1-14., [PubMed]

Sentences

No. Mutations Sentence Comment

59 ABCB1 p.Gly185Val A single amino acid substitution, Gly185Val, in the human ABCB1 protein was found to cause an altered pattern of drug resistance in cell lines transfected with the ABCB1 cDNA carrying this mutation.21) It is suggested that the amino acid at position 185 is involved in colchicines and verapamil but not in vinblastine bindingWtransport. Login to comment 78 ABCB1 p.Gly185Val ABCB1 p.Ser400Asn ABCB1 p.Asn21Asp ABCB1 p.Phe103Leu ABCB1 p.Met986Val ABCB1 p.Asn44Ser For this purpose, the cDNA of ABCB1 was cloned from the human liver cDNA library, and several variant forms (i.e., N21D, N44S, F103L, G185V, S400N, A893S, A893T, M986V) were prepared by site-directed mutagenesis (see Fig. 2A for primers). Login to comment 86 ABCB1 p.Gly185Val ABCB1 p.Ser400Asn ABCB1 p.Asn21Asp ABCB1 p.Phe103Leu ABCB1 p.Met986Val ABCB1 p.Asn44Ser The variant forms (i.e., N21D, N44S, F103L, G185V, S400N, A893S, A893T, M986V) exhibited the verapamil-enhanced ATPase activity, as did the wild type of ABCB1. Login to comment 89 ABCB1 p.Gly185Val ABCB1 p.Asn21Asp The variant G185V (acquired mutation) was found to have the highest Vmax value, which was followed by N21D (Table 2). Login to comment 105 ABCB1 p.Gly185Val ABCB1 p.Ser400Asn ABCB1 p.Asn21Asp ABCB1 p.Phe103Leu ABCB1 p.Met986Val ABCB1 p.Asn44Ser Kinetic parameters of the wild type and SNP variants of ABCB1 Variant Km Vmax (mM) (nmolWminWmg protein) Wild type 2.190±0.150 13.14±1.95 N21D 0.502±0.126 45.26±11.33 N44S 0.580±0.148 31.03±4.65 F103L 1.100±0.078 36.34±8.33 G185V 0.831±0.102 56.76±6.76 S400N 0.327±0.025 13.74±2.08 A893S 0.441±0.042 17.24±6.72 A893T 0.904±0.244 10.77±1.35 M986V 0.419±0.062 22.69±6.84 The wild type and variants of ABCB1 were then expressed it in Sf9 cells using the pFASTBAC1 vector and recombinant baculoviruses. Login to comment