ABCMdb

ABCA4 p.Lys1978Ala

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Publications

PMID: 16533065 [PubMed] Biswas-Fiss EE et al: "Interaction of the nucleotide binding domains and regulation of the ATPase activity of the human retina specific ABC transporter, ABCR."

No. Sentence Comment

74 The following complementary oligonucleotides were used as mutagenic primers to generate the mutant NBD1 and NBD2 proteins: NBD1Pro940Arg (5'-GGT AAA GAT TTT TGA GCG CTG TGG CCG GCC AGC TG-3'), NBD1 Walker A, K969A, T970A (5'-CAA TGG AGC TGG GGC AGC CAC CAC CTT GTC CAT CC-3'), NBD2 Walker A, K1978A, T1979A (5'-GAA TGG TGC CGG CGC AGC AAC CAC ATT CAA GAT GC-3'), and NBD2 R2038W (5'-CTT TAC CTT TAT GCC AGG CTT CGA GGT GTA CCA GC-3'). Login to comment
199 The affinity of interaction decreased under all conditions examined, as determined by comparison of the Kd values (Table Table 2: Modulation of ATPase Activities of NBD1‚NBD2 Complexes by Site-Specific Mutations NBD1‚NBD2 mutation domain NBD1‚NBD2 ATPasea (pmol/min) NBD1‚NBD2 ATPaseexptl (pmol/min) NBD1‚NBD2 ATPasecalcd b (pmol/min) motif/disease NBD1mut‚NBD2wt K969A, T970A NBD1 1.9 ( 0.4/5.3 ( 0.1 5.1 ( 0.1 7.2 synthetic NBD1wt‚NBD2mut K1978A, 1979A NBD2 3.1 ( 0.2/1.6 ( 0.3 1.5 ( 0.2 4.7 synthetic NBD1mut‚NBD2wt P940R NBD1 1.9 ( 0.3/5.2 ( 0.2 4.3 ( 0.1 7.1 AMD NBD1wt‚NBD2mut R2038W NBD2 3.1 ( 0.1/2.0 ( 0.2 4.4 ( 0.2 5.1 STGD NBD1wt‚NBD2wt wild type N/A 3.0 ( 0.3/5.4 ( 0.1 5.30 ( 0.3 8.4 none a Experimental ATPase activity corresponding to 6 pmol of the individual domains, NBD1 and NBD2, respectively. Login to comment
218 On the other hand, the ratio of the experimental versus theoretical ATPase activities of the complex harboring NBD2 Walker A mutations, K1978A, T1979A, in the NBD2 domain was only 0.32, which represents a 68% difference in the experimental versus the theoretical activity, as compared to the 36% difference observed for the wild-type complex. Login to comment
225 The ATPase assay was carried out as described under Experimental Procedures at 37 °C for 60 min. A theoretical curve is shown, indicating the results expected if the activity of NBD1K969A,T970A‚ NBD2 were additive (]) of that obtained for the individual domains. Each point represents the average of three independent experiments. FIGURE 7: Effect of NBD2 Walker A mutations K1978A, T1979A on modulation of ATP hydrolysis of the NBD1‚NBD2 protein complex. Protein titration of the ATPase activity of purified NBD2K1978A,T1979A (∆), NBD2WT (2), NBD1WT (0) alone, and the NBD1‚NBD2K1978A,T1979A complex (O). Login to comment