ABCMdb

PMID: 10551832

Liu LX, Janvier K, Berteaux-Lecellier V, Cartier N, Benarous R, Aubourg P
Homo- and heterodimerization of peroxisomal ATP-binding cassette half-transporters.
J Biol Chem. 1999 Nov 12;274(46):32738-43., [PubMed]

Sentences

No. Mutations Sentence Comment

45 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln ALD point mutations (R389H, R401Q, P484R, and R591Q) were individually introduced into pGBT- and pLEX-hALDPc by site-directed mutagenesis using "overlap extension PCR" with Pfu polymerase (Stratagene) and appropriate primers (25). Login to comment 46 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln ALD point mutations (R389H, R401Q, P484R, and R591Q) were individually introduced into pGBT- and pLEX-hALDPc by site-directed mutagenesis using "overlap extension PCR" with Pfu polymerase (Stratagene) and appropriate primers (25). Login to comment 47 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln ALD point mutations (R389H, R401Q, P484R, and R591Q) were individually introduced into pGBT- and pLEX-hALDPc by site-directed mutagenesis using "overlap extension PCR" with Pfu polymerase (Stratagene) and appropriate primers (25). Login to comment 70 ABCD1 p.Arg389His ABCD1 p.Arg591Gln ABCD2 p.Pro484Arg These mutations (R389H, R401Q, P484R, and R591Q) were generated as described above and tested in two-hybrid assays. Login to comment 71 ABCD1 p.Arg389His ABCD1 p.Arg591Gln ABCD2 p.Pro484Arg ABCD2 p.Pro484Arg These mutations (R389H, R401Q, P484R, and R591Q) were generated as described above and tested in two-hybrid assays. Login to comment 72 ABCD1 p.Arg389His ABCD1 p.Arg389His ABCD1 p.Arg591Gln ABCD2 p.Pro484Arg ABCD2 p.Pro484Arg These mutations (R389H, R401Q, P484R, and R591Q) were generated as described above and tested in two-hybrid assays. Login to comment 73 ABCD1 p.Arg389His ABCD1 p.Arg591Gln ABCD2 p.Pro484Arg ABCD2 p.Pro484Arg The P484R mutation leads to a decreased amount of ALDP in patient fibroblasts,3 whereas the three other mutations have no effect on ALDP stability in vivo (28-31). Login to comment 74 ABCD1 p.Arg389His ABCD1 p.Arg591Gln ABCD2 p.Pro484Arg Our results show that the mutations R389H and R401Q had no effect on the interactions of hALDPc with itself (Fig. 2A, rows 1, 3, and 5), mALDRPc (Fig. 2B, rows 1, 3, and 5), or hPMP70c (Fig. 2C, rows 1, 3, and 5). Login to comment 75 ABCD1 p.Arg591Gln ABCD2 p.Pro484Arg In contrast, the P484R and R591Q mutations 2 L. X. Liu, K. Janvier, V. Berteaux-Lecellier, N. Cartier, R. Benarous, and P. Aubourg, unpublished results. Login to comment 88 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln B and C, interactions of wild type and mutated (R389H, R401Q, P484R, and R591Q) ALDPc with mALDRPc and hPMP70c. Login to comment 89 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln B and C, interactions of wild type and mutated (R389H, R401Q, P484R, and R591Q) ALDPc with mALDRPc and hPMP70c. Login to comment 90 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln B and C, interactions of wild type and mutated (R389H, R401Q, P484R, and R591Q) ALDPc with mALDRPc and hPMP70c. Login to comment 93 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln A, HF7c yeast strains expressing wild type or mutated (R389H, R401Q, P484R, and R591Q) ALDPc fused to Gal4-BD were analyzed for histidine auxotrophy (left panel, medium with histidine; right panel, medium without histidine). Login to comment 94 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln A, HF7c yeast strains expressing wild type or mutated (R389H, R401Q, P484R, and R591Q) ALDPc fused to Gal4-BD were analyzed for histidine auxotrophy (left panel, medium with histidine; right panel, medium without histidine). Login to comment 95 ABCD1 p.Arg401Gln ABCD1 p.Pro484Arg ABCD1 p.Arg389His ABCD1 p.Arg591Gln A, HF7c yeast strains expressing wild type or mutated (R389H, R401Q, P484R, and R591Q) ALDPc fused to Gal4-BD were analyzed for histidine auxotrophy (left panel, medium with histidine; right panel, medium without histidine). Login to comment 147 ABCD2 p.Pro484Arg Interestingly, the P484R mutation results in an unstable protein in vivo. Login to comment 148 ABCD2 p.Pro484Arg Interestingly, the P484R mutation results in an unstable protein in vivo. Login to comment 149 ABCD2 p.Pro484Arg Interestingly, the P484R mutation results in an unstable protein in vivo. Login to comment 150 ABCD1 p.Arg591Gln In contrast, the R591Q disease mutation, which alters the dimerization of ALDP in the yeast two-hybrid assays, does not lead to ALDP unstability in vivo. Login to comment 151 ABCD1 p.Arg591Gln In contrast, the R591Q disease mutation, which alters the dimerization of ALDP in the yeast two-hybrid assays, does not lead to ALDP unstability in vivo. Login to comment 152 ABCD1 p.Pro560Leu ABCD1 p.Arg518Trp ABCD1 p.Arg591Gln In contrast, the R591Q disease mutation, which alters the dimerization of ALDP in the yeast two-hybrid assays, does not lead to ALDP unstability in vivo. Login to comment 153 ABCD1 p.Pro560Leu ABCD1 p.Arg518Trp This is supported by the observation that several ALDP missense mutations (in particular R518W and P560L) can lead to either a normal, decreased level or absence of ALDP in ALD fibroblasts.4 The specific carboxyl-terminal subdomain region of ALDP responsible for dimerization could not be defined. Login to comment 154 ABCD1 p.Pro560Leu ABCD1 p.Arg518Trp This is supported by the observation that several ALDP missense mutations (in particular R518W and P560L) can lead to either a normal, decreased level or absence of ALDP in ALD fibroblasts.4 The specific carboxyl-terminal subdomain region of ALDP responsible for dimerization could not be defined. Login to comment