ABCMdb

ABCD3 p.Ile70Ala

None has been submitted yet.

Publications

PMID: 17761678 [PubMed] Kashiwayama Y et al: "Hydrophobic regions adjacent to transmembrane domains 1 and 5 are important for the targeting of the 70-kDa peroxisomal membrane protein."

No. Sentence Comment

88 The lysate was cen- TABLE 2 Oligonucleotide primer sequences used for the generation of mutant PMP70 constructs Conctruct name Forward primer (5؅ 3 3؅) K28A/R29A GCTCTGCCTGCTCCACGCGGCGCGCCGCGCCCTCG R30A/R31A CCTGCTCCACGCGGCGGCCGCCGCCCTCGGCCTGCACG K38A/K39A CCTCGGCCTGCACGGTGCGGCAAGTGGAAAACCACCATTAC P76A/R77A CAGATTCTGAAAATCATGGTCGCTGCAACATTTTGTAAAGAGACAGG K72A GGCTCATACAGATTCTGGCAATCATGGTCGCTGCAAC R66A GGACAAGGTGTTTTTCTCAGCGCTCATACAGATTCTGGC K61A CGAGCTGTGGTGGACGCGGTGTTTTTCTCAGC K53A/K54A/R56A CAATGAGAAAGAGGGGGCAGCAGAGGCAGCTGTGGTGGACGCGG K123A/R124A GGTCGTAGCAGGAAAGATTTCGCGGCATACTTACTCAACTTCATCG R119A/K120A GGTATCATTGGTCGTAGCGCGGCAGATTTCGCGGCATACTTACTC R117A GTGGTATCATTGGTGCTAGCGCGGCAGATTTCGCG L21A/L22A/L23A GTGCCGCGTTCGCTGCTGCTTGCCTGCTCCAC L21Q/L22Q/L23Q GCTGGTGCCGCGTTCCAGCAGCAGTGCCTGCTCCACAAGC I70A/L71A CTCAAGGCTCATACAGGCTGCGAAAATCATGGTCCCTAGAAC I70N/L71Q CTCAAGGCTCATACAGAATCAGAAAATCATGGTCCCTAGAAC I307N/L308Q GGTGGAACACCTACATAATTTCAATCAGTTTCGGTTTTCAATGGGC Peroxisome Targeting Signal of PMP70 NOVEMBER 16, 2007•VOLUME 282•NUMBER 46 JOURNAL OF BIOLOGICAL CHEMISTRY 33833 trifuged at 20,000 ϫ g for 30 min and the His-Pex19p in the supernatant was immediately applied to 10 ml of TALON Metal affinity resin (Clontech) equilibrated with the lysis buffer. Login to comment
176 Furthermore, only 15% of the fluorescence of PMP70(AA.224-375)- GFP coincided with that of peroxisomes, which was comparable PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144 I70N/L71Q)-GFP Not detected Not detected PMP70(AA.1-144 I70A/L71A)-GFP PMP70(AA.1-144 L21A/L22A/L23A)-GFP (A) 45.0 31.0 kDa 45.0 31.0 kDa 01enoN µM MG132 (B) PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144 I70N/L71Q)-GFP PMP70(AA.1-659 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-659 I70N/L71Q)-GFP (C) control PMP70(AA.1-144I70N/L71Q)-GFP PMP70(AA.1-144L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144)-GFP control GFP PMP70(AA.1-144I70A/L71A)-GFP PMP70(AA.1-144I70N/L71Q)-GFP PMP70(AA.1-144L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144)-GFP FIGURE 4. Login to comment
178 A, PMP70(AA.1-144 L21A/L22A/L23A)-GFP, PMP70(AA.1-144 I70A/L71A)-GFP, PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP, and PMP70(AA.1-144 I70N/L71Q)-GFP were expressed in CHO cells. The subcellular distribution of the fusion proteins was compared with the localization of the endogenous PMP70 detected by immunofluorescence staining with anti-PMP70 COOH-terminal antibody. The peroxisomal staining pattern is shown on the left, EGFP fluorescence in the center, and a superimposition of both stains on the right. Login to comment
236 2) PMP70(AA.1-144 L21A/L22A/ L23A)-GFP and PMP70(AA.1-144 I70A/L71A)-GFP, which possess a hydrophobic property similar to that of PMP70(AA.1-144)-GFP, were localized to peroxisomes. Login to comment
237 On the other hand, both PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP and PMP70(AA.1-144 I70N/L71Q)- GFP were degraded by proteasomes and did not show any peroxisomal localization even in the presence of proteasome inhibitor (Fig. 4). Login to comment
89 The lysate was cen- TABLE 2 Oligonucleotide primer sequences used for the generation of mutant PMP70 constructs Conctruct name Forward primer (5d15; 3 3d15;) K28A/R29A GCTCTGCCTGCTCCACGCGGCGCGCCGCGCCCTCG R30A/R31A CCTGCTCCACGCGGCGGCCGCCGCCCTCGGCCTGCACG K38A/K39A CCTCGGCCTGCACGGTGCGGCAAGTGGAAAACCACCATTAC P76A/R77A CAGATTCTGAAAATCATGGTCGCTGCAACATTTTGTAAAGAGACAGG K72A GGCTCATACAGATTCTGGCAATCATGGTCGCTGCAAC R66A GGACAAGGTGTTTTTCTCAGCGCTCATACAGATTCTGGC K61A CGAGCTGTGGTGGACGCGGTGTTTTTCTCAGC K53A/K54A/R56A CAATGAGAAAGAGGGGGCAGCAGAGGCAGCTGTGGTGGACGCGG K123A/R124A GGTCGTAGCAGGAAAGATTTCGCGGCATACTTACTCAACTTCATCG R119A/K120A GGTATCATTGGTCGTAGCGCGGCAGATTTCGCGGCATACTTACTC R117A GTGGTATCATTGGTGCTAGCGCGGCAGATTTCGCG L21A/L22A/L23A GTGCCGCGTTCGCTGCTGCTTGCCTGCTCCAC L21Q/L22Q/L23Q GCTGGTGCCGCGTTCCAGCAGCAGTGCCTGCTCCACAAGC I70A/L71A CTCAAGGCTCATACAGGCTGCGAAAATCATGGTCCCTAGAAC I70N/L71Q CTCAAGGCTCATACAGAATCAGAAAATCATGGTCCCTAGAAC I307N/L308Q GGTGGAACACCTACATAATTTCAATCAGTTTCGGTTTTCAATGGGC Peroxisome Targeting Signal of PMP70 NOVEMBER 16, 2007ߦVOLUME 282ߦNUMBER 46 JOURNAL OF BIOLOGICAL CHEMISTRY 33833 trifuged at 20,000 afb; g for 30 min and the His-Pex19p in the supernatant was immediately applied to 10 ml of TALON Metal affinity resin (Clontech) equilibrated with the lysis buffer. Login to comment
177 Furthermore, only 15% of the fluorescence of PMP70(AA.224-375)- GFP coincided with that of peroxisomes, which was comparable PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144 I70N/L71Q)-GFP Not detected Not detected PMP70(AA.1-144 I70A/L71A)-GFP PMP70(AA.1-144 L21A/L22A/L23A)-GFP (A) 45.0 31.0 kDa 45.0 31.0 kDa 0 1 e n o N &#b5;M MG132 (B) PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144 I70N/L71Q)-GFP PMP70(AA.1-659 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-659 I70N/L71Q)-GFP (C) control PMP70(AA.1-144 I70N/L71Q)-GFP PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144)-GFP control GFP PMP70(AA.1-144 I70A/L71A)-GFP PMP70(AA.1-144 I70N/L71Q)-GFP PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP PMP70(AA.1-144)-GFP FIGURE 4. Login to comment
179 A, PMP70(AA.1-144 L21A/L22A/L23A)-GFP, PMP70(AA.1-144 I70A/L71A)-GFP, PMP70(AA.1-144 L21Q/L22Q/L23Q)-GFP, and PMP70(AA.1-144 I70N/L71Q)-GFP were expressed in CHO cells. The subcellular distribution of the fusion proteins was compared with the localization of the endogenous PMP70 detected by immunofluorescence staining with anti-PMP70 COOH-terminal antibody. The peroxisomal staining pattern is shown on the left, EGFP fluorescence in the center, and a superimposition of both stains on the right. Login to comment