ABCMdb

ABCD3 p.Arg31Ala

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
128 As shown in Fig. 3B, PMP70(AA.1-144 K28A/R29A/R30A/ R31A/K38A/K39A)-GFP,PMP70(AA.1-144K53A/K54A/R56A/ K61A/R66A/K72A/R77A)-GFP, and PMP70(AA.1-144 R117A/ R119A/K120A/K123A/R124A)-GFP were localized to peroxisomes, and the targeting efficiency of each mutant protein was almost the same as that of PMP70(AA.1-144)-GFP. Login to comment
129 Under the same condition, PMP70(AA.1-144 L21Q/L22Q/L23Q)- GFP and PMP70(AA.1-144 I70N/L71Q)-GFP were not localized to peroxisomes as negative controls (see Fig. 4). Login to comment
154 PMP70(AA.1-659 I70N/L71Q) existed in a soluble form in the presence of His-Pex19p and still interacted with His-Pex19p at almost the same level as wild type PMP70 did, but the mutant PMP70 was mislocalized to endoplasmic reticulum-like structures, as (B) (A) PMP70(AA.1-144 K28A/R29A/R30A/R31A/K38A/K39A)-GFP PMP70(AA.1-144 K53A/K54A/R56A/K61A/R66A/K72A/P76A/R77A)-GFP PMP70(AA.1-144 R117A/R119A/K120A/K123A/R124A)-GFP MAAFSKYLT-----------ARNSS-LAGAAFLL--LCLLHKRRRALGLHGKKS-- ------ GKPPLQNNEKE MPVLSRPRP----W-RGNTLKRTAVLLALAAYGAHKVYPLVRQCL-APARGLQAPAGEPTQEASGV------- MTHMLNAAADRVKWTRSSAA CLVA-AAYALKTLYPIIGKRLKQSGHGKKKAAAYPAAENTEILHCTETIKRAA GKKERAVVDKVFFSRLIQILKIMVPRTFCKETGYLVLIAVMLVSRTYCDVWMIQNGTLIESGIIGRSRKDFKR -AAAKAGMNRVFLQRLLWLLRLLFPRVLCRETGLLALHSAALVSRTFLSVYVARLDGRLARCIARKDPRAFGW CEKPSPGVNADFFKQLLELRKILFPKLVTTETGWLCLHSVALISRTFLSIYVAGLDGKIVKSIVEKKPRTFII ++++ ++ ++ + +++++ + + ++ 1 1 1 52 61 73 51 60 72 124 132 145 PMP70 ALDP ALDRP PMP70 ALDP ALDRP TMD1 FIGURE3.BasicaminoacidresiduesintheNH2-terminalregionofPMP70arenotnecessaryforthetargetingofPMP70.A,alignmentoftheNH2-terminal region of human peroxisomal ABC transporters. 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
155 PMP70(AA.1-659 I70N/L71Q) existed in a soluble form in the presence of His-Pex19p and still interacted with His-Pex19p at almost the same level as wild type PMP70 did, but the mutant PMP70 was mislocalized to endoplasmic reticulum-like structures, as (B) (A) PMP70(AA.1-144 K28A/R29A/R30A/R31A/K38A/K39A)-GFP PMP70(AA.1-144 K53A/K54A/R56A/K61A/R66A/K72A/P76A/R77A)-GFP PMP70(AA.1-144 R117A/R119A/K120A/K123A/R124A)-GFP MAAFSKYLT-----------ARNSS-LAGAAFLL--LCLLHKRRRALGLHGKKS-- ----- - GKPPLQNNEKE MPVLSRPRP----W-RGNTLKRTAVLLALAAYGAHKVYPLVRQCL-APARGLQAPAGEPTQEASGV------- MTHMLNAAADRVKWTRSSAA CLVA-AAYALKTLYPIIGKRLKQSGHGKKKAAAYPAAENTEILHCTETI KRAA GKKERAVVDKVFFSRLIQILKIMVPRTFCKETGYLVLIAVMLVSRTYCDVWMIQNGTLIESGIIGRSRKDFKR -AAAKAGMNRVFLQRLLWLLRLLFPRVLCRETGLLALHSAALVSRTFLSVYVARLDGRLARCIARKDPRAFGW CEKPSPGVNADFFKQLLELRKILFPKLVTTETGWLCLHSVALISRTFLSIYVAGLDGKIVKSIVEKKPRTFII ++++ ++ ++ + + + + + + + + ++ 1 1 1 52 61 73 51 60 72 124 132 145 PMP70 ALDP ALDRP PMP70 ALDP ALDRP TMD1 FIGURE3.BasicaminoacidresiduesintheNH2-terminalregionofPMP70arenotnecessaryforthetargetingofPMP70.A,alignmentoftheNH2-terminal region of human peroxisomal ABC transporters. Login to comment