ABCMdb

ABCA1 p.Ser1255Ala

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Publications

PMID: 15218032 [PubMed] Roosbeek S et al: "Phosphorylation by protein kinase CK2 modulates the activity of the ATP binding cassette A1 transporter."

No. Sentence Comment

133 In both systems we engineered several mutants designed at preventing phosphorylation by mutation either of the target threonines and serine, T1242A, T1243A, T1242A/ T1243A, S1255A, or of the downstream cluster of acidic residues, E1245Q, E1246Q, E1245Q/E1246Q. Login to comment
144 Analysis of the S1255A mutant demonstrates the loss of one phosphorylation site in the Val-1251-Arg-1272 tryptic peptide (Table I). Login to comment
159 Compared with WT ABCA1, set as 100%, the PE and PS content in the outer leaflet of the plasma membrane increased up to 130 Ϯ 14 and 123 Ϯ 8% for the T1242A mutant, 154 Ϯ 15 and 148 Ϯ 11% for the T1243A mutant, 170 Ϯ 15 and 207 Ϯ 16% for the T1242A/T1243A mutant, 141 Ϯ 11 and 136 Ϯ 10% for the S1255A mutant, and 141 Ϯ 13 and 120 Ϯ 9% for the E1245Q/E1246Q mutant, respectively (Fig. 4). Login to comment
162 The effect of the specific CK2 inhibitors 4,5,6,7-tetrabromo- benzotriazole and apigenin on the flippase activity in Hek-293 Tet-Off cells transfected with WT ABCA1 and with the T1242A, T1243A, T1242A/T1243A, S1255A, and T1242D/ T1243D mutants was consistent with the above data. Login to comment
176 TABLE I Mass spectrometric analysis of NBD1ϩR1 phosphorylation NBD1ϩR1 Tryptic peptide Molecular mass Molecular mass after beta-elimination Phosphorylated residues Da Da WT Leu-1229-Arg-1269 4542.5 4249.5 Thr-1242, Thr-1243, Ser-1255 T1242A Leu-1229-Lys-1250 2483.6 2385.9 Thr-1243 T1242A Val-1251-Arg-1272 2411.6 2313.7 Ser-1255 T1243A Leu-1229-Lys-1250 2483.1 2385.2 Thr-1242 T1243A Val-1251-Arg-1272 2411.2 2313.2 Ser-1255 T1242A/T1243A Leu-1229-Lys-1250 2356.1 2356.1 T1242A/T1243A Val-1251-Arg-1272 2411.6 2313.7 Ser-1255 S1255A Val-1251-Arg-1272 2297.7 2297.7 compared with WT ABCA1. Login to comment
177 This increase was more pronounced for the T1243A mutation either alone or in combination with the T1242A mutation, whereas the effect of the T1242A and S1255A mutants was similar. Login to comment
196 of the S1255A mutant on lipid efflux was comparable with the single threonine mutants. Login to comment
213 This was then confirmed by the engineering of the S1255A mutant, whose activities are comparable with those of the T1242A mutant. Login to comment
131 In both systems we engineered several mutants designed at preventing phosphorylation by mutation either of the target threonines and serine, T1242A, T1243A, T1242A/ T1243A, S1255A, or of the downstream cluster of acidic residues, E1245Q, E1246Q, E1245Q/E1246Q. Login to comment
142 Analysis of the S1255A mutant demonstrates the loss of one phosphorylation site in the Val-1251-Arg-1272 tryptic peptide (Table I). Login to comment
157 Compared with WT ABCA1, set as 100%, the PE and PS content in the outer leaflet of the plasma membrane increased up to 130 afe; 14 and 123 afe; 8% for the T1242A mutant, 154 afe; 15 and 148 afe; 11% for the T1243A mutant, 170 afe; 15 and 207 afe; 16% for the T1242A/T1243A mutant, 141 afe; 11 and 136 afe; 10% for the S1255A mutant, and 141 afe; 13 and 120 afe; 9% for the E1245Q/E1246Q mutant, respectively (Fig. 4). Login to comment
160 The effect of the specific CK2 inhibitors 4,5,6,7-tetrabromo- benzotriazole and apigenin on the flippase activity in Hek-293 Tet-Off cells transfected with WT ABCA1 and with the T1242A, T1243A, T1242A/T1243A, S1255A, and T1242D/ T1243D mutants was consistent with the above data. Login to comment
174 TABLE I Mass spectrometric analysis of NBD1af9;R1 phosphorylation NBD1af9;R1 Tryptic peptide Molecular mass Molecular mass after beta-elimination Phosphorylated residues Da Da WT Leu-1229-Arg-1269 4542.5 4249.5 Thr-1242, Thr-1243, Ser-1255 T1242A Leu-1229-Lys-1250 2483.6 2385.9 Thr-1243 T1242A Val-1251-Arg-1272 2411.6 2313.7 Ser-1255 T1243A Leu-1229-Lys-1250 2483.1 2385.2 Thr-1242 T1243A Val-1251-Arg-1272 2411.2 2313.2 Ser-1255 T1242A/T1243A Leu-1229-Lys-1250 2356.1 2356.1 T1242A/T1243A Val-1251-Arg-1272 2411.6 2313.7 Ser-1255 S1255A Val-1251-Arg-1272 2297.7 2297.7 Protein Kinase CK2 Phosphorylation Modulates ABCA1 Activity compared with WT ABCA1. Login to comment
175 This increase was more pronounced for the T1243A mutation either alone or in combination with the T1242A mutation, whereas the effect of the T1242A and S1255A mutants was similar. Login to comment
194 of the S1255A mutant on lipid efflux was comparable with the single threonine mutants. Login to comment
211 This was then confirmed by the engineering of the S1255A mutant, whose activities are comparable with those of the T1242A mutant. Login to comment