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  • 99
    Thermo Fisher antibody library
    Antibody Library, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore dtt
    Dtt, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 36382 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare dtt
    Dtt, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 18540 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Syntaxin 3t3 l1 adipocytes
    3t3 L1 Adipocytes, supplied by Syntaxin, used in various techniques. Bioz Stars score: 92/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad protean ief cell
    Protean Ief Cell, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 5637 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Syntaxin vamp4
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Vamp4, supplied by Syntaxin, used in various techniques. Bioz Stars score: 92/100, based on 550 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GE Healthcare ipgphor ii
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Ipgphor Ii, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 661 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad rehydration buffer
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Rehydration Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1818 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher revertaid reverse transcriptase
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Revertaid Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    GE Healthcare rehydration buffer
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Rehydration Buffer, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 98/100, based on 3142 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    GE Healthcare destreak reagent
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Destreak Reagent, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 691 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GE Healthcare immobiline drystrip
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Immobiline Drystrip, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 1325 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    GE Healthcare ipg strips
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Ipg Strips, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 95/100, based on 5984 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher revertaid h minus first strand cdna synthesis kit
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Revertaid H Minus First Strand Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9555 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GE Healthcare 2 d quant kit
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    2 D Quant Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 4658 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    2 d quant kit - by Bioz Stars, 2020-09
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    99
    Thermo Fisher lysis buffer
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 41889 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore tris
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Tris, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 38479 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Burkert beck am
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Beck Am, supplied by Burkert, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore sodium acetate
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Sodium Acetate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5584 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore pluronic f 127
    Expression of PACS-1 admut causes mislocalization of <t>VAMP4.</t> AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.
    Pluronic F 127, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2302 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression of PACS-1 admut causes mislocalization of VAMP4. AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.

    Journal: EMBO Reports

    Article Title: AP-1 recruitment to VAMP4 is modulated by phosphorylation-dependent binding of PACS-1

    doi: 10.1038/sj.embor.7400018

    Figure Lengend Snippet: Expression of PACS-1 admut causes mislocalization of VAMP4. AtT20 cells were transfected with Flag-VAMP4 and after 60 h infected with either ( A ) wt, ( B ) HA-PACS-1 or ( C ) PACS-1 admut vaccinia virus. Flag-VAMP4 panels (green, detected with M2 ab) show the localization of VAMP4, ACTH panels (red, detected with anti-POMC ab) show the localization of POMC and ACTH, and MERGE panels show the merge of Flag and ACTH. Expression of PACS-1 admut (see the arrowhead in ( C )) but not PACS-1 (see arrowheads in ( A , B )) causes mislocalization of Flag-VAMP4 to the ACTH-positive MSGs in the tips of the cells. The insets in ( C ) show double labelling of VAMP4 and ACTH in two other tips.

    Article Snippet: Since VAMP4 contains an acidic dileucine motif similar to CI-MPR , we tested for interaction between VAMP4 and the VHS domains of the GGAs.

    Techniques: Expressing, Transfection, Infection

    Phosphoserine 30 modulates AP-1 binding to VAMP4 via PACS-1. ( A ) AtT20 cells were infected with either wt or HA-PACS-1 vaccinia virus and used for immunoprecipitation with anti-VAMP4 antibody. HA-PACS-1 can be detected in the VAMP4 immunoprecipitates. ( B ) GST-VAMP4 was incubated with or without CK2, bound to glutathione beads and incubated with PACS-1. PACS-1 binding to wt or the LL/VV VAMP4 is enhanced by the presence of CK2; the S30D mutant shows a CK2-independent binding comparable to phosphorylated VAMP4. The lower panel shows quantitation of the PACS-1 binding to wt VAMP4 and mutants in the presence or absence of CK2. ( C ) Formation of the ternary VAMP4/PACS-1/AP-1 complex using GST-VAMP4, PACS-1 and purified AP-1. Addition of PACS-1 protein to the VAMP4 LL/VV S30D protein enhances binding of AP-1 compared to VAMP4 LL/VV S30A protein. Quantitation (lower panel) of AP-1 binding to the VAMP4/PACS-1 complex shows a PACS-1-dependent increase.

    Journal: EMBO Reports

    Article Title: AP-1 recruitment to VAMP4 is modulated by phosphorylation-dependent binding of PACS-1

    doi: 10.1038/sj.embor.7400018

    Figure Lengend Snippet: Phosphoserine 30 modulates AP-1 binding to VAMP4 via PACS-1. ( A ) AtT20 cells were infected with either wt or HA-PACS-1 vaccinia virus and used for immunoprecipitation with anti-VAMP4 antibody. HA-PACS-1 can be detected in the VAMP4 immunoprecipitates. ( B ) GST-VAMP4 was incubated with or without CK2, bound to glutathione beads and incubated with PACS-1. PACS-1 binding to wt or the LL/VV VAMP4 is enhanced by the presence of CK2; the S30D mutant shows a CK2-independent binding comparable to phosphorylated VAMP4. The lower panel shows quantitation of the PACS-1 binding to wt VAMP4 and mutants in the presence or absence of CK2. ( C ) Formation of the ternary VAMP4/PACS-1/AP-1 complex using GST-VAMP4, PACS-1 and purified AP-1. Addition of PACS-1 protein to the VAMP4 LL/VV S30D protein enhances binding of AP-1 compared to VAMP4 LL/VV S30A protein. Quantitation (lower panel) of AP-1 binding to the VAMP4/PACS-1 complex shows a PACS-1-dependent increase.

    Article Snippet: Since VAMP4 contains an acidic dileucine motif similar to CI-MPR , we tested for interaction between VAMP4 and the VHS domains of the GGAs.

    Techniques: Binding Assay, Infection, Immunoprecipitation, Incubation, Mutagenesis, Quantitation Assay, Purification, Protein Quantitation

    Sorting of VAMP4 in AtT20 cells requires both AP-1 and PACS-1 binding sites. The localization of transfected Flag-tagged ( A ) wt VAMP4, ( B ) VAMP4 LL/VV, ( C ) VAMP4 S30A or ( D ) VAMP4 LL/VV S30A was compared to endogenous ACTH in AtT20 cells. The left panels show the Flag tag (green, M2 ab), the centre panels show ACTH (red, POMC ab) and the right panels show the MERGE of the two images. The wt, LL/VV and S30A proteins remain predominantly in a perinuclear region, and are not found in the tips (arrowheads in ( A – C )) with ACTH. Mutation of both the dileucine motif and the PACS-1 binding motif results in mislocalization of VAMP4 to ACTH-positive MSGs found in the tips of the cells (arrowhead in ( D )). The inset in ( C ) MERGE shows double labelling of VAMP4 and ACTH in another tip.

    Journal: EMBO Reports

    Article Title: AP-1 recruitment to VAMP4 is modulated by phosphorylation-dependent binding of PACS-1

    doi: 10.1038/sj.embor.7400018

    Figure Lengend Snippet: Sorting of VAMP4 in AtT20 cells requires both AP-1 and PACS-1 binding sites. The localization of transfected Flag-tagged ( A ) wt VAMP4, ( B ) VAMP4 LL/VV, ( C ) VAMP4 S30A or ( D ) VAMP4 LL/VV S30A was compared to endogenous ACTH in AtT20 cells. The left panels show the Flag tag (green, M2 ab), the centre panels show ACTH (red, POMC ab) and the right panels show the MERGE of the two images. The wt, LL/VV and S30A proteins remain predominantly in a perinuclear region, and are not found in the tips (arrowheads in ( A – C )) with ACTH. Mutation of both the dileucine motif and the PACS-1 binding motif results in mislocalization of VAMP4 to ACTH-positive MSGs found in the tips of the cells (arrowhead in ( D )). The inset in ( C ) MERGE shows double labelling of VAMP4 and ACTH in another tip.

    Article Snippet: Since VAMP4 contains an acidic dileucine motif similar to CI-MPR , we tested for interaction between VAMP4 and the VHS domains of the GGAs.

    Techniques: Binding Assay, Transfection, FLAG-tag, Mutagenesis

    Binding of VAMP4 to AP-1 is modulated by CK2. ( A ) VAMP4 contains a dileucine motif surrounded by two acidic clusters (boxed). ( B ) GST-syntaxin 6 or -VAMP4 were pretreated with or without CK2, bound to glutathione beads and incubated with BAMC. AP-1 binds GST-VAMP4, but not syntaxin 6, and binding is enhanced by CK2 phosphorylation. AP-2 does not bind either syntaxin 6 or VAMP4. ( C ) Immobilized his-GGA-VHS domains were incubated with GST-CI-MPR or -VAMP4. GGA1 bound to the CI-MPR, while no binding of GGA1, 2 or 3-VHS to VAMP4 was detected (data not shown). ( D ) Yeast was co-transformed with VAMP4 2–60 and the GGA1–3-VHS domain, and histidine-dependent growth was assessed. The GGAs do not bind to VAMP4 2–60 , while specific GGA interaction with CI-MPR was detected.

    Journal: EMBO Reports

    Article Title: AP-1 recruitment to VAMP4 is modulated by phosphorylation-dependent binding of PACS-1

    doi: 10.1038/sj.embor.7400018

    Figure Lengend Snippet: Binding of VAMP4 to AP-1 is modulated by CK2. ( A ) VAMP4 contains a dileucine motif surrounded by two acidic clusters (boxed). ( B ) GST-syntaxin 6 or -VAMP4 were pretreated with or without CK2, bound to glutathione beads and incubated with BAMC. AP-1 binds GST-VAMP4, but not syntaxin 6, and binding is enhanced by CK2 phosphorylation. AP-2 does not bind either syntaxin 6 or VAMP4. ( C ) Immobilized his-GGA-VHS domains were incubated with GST-CI-MPR or -VAMP4. GGA1 bound to the CI-MPR, while no binding of GGA1, 2 or 3-VHS to VAMP4 was detected (data not shown). ( D ) Yeast was co-transformed with VAMP4 2–60 and the GGA1–3-VHS domain, and histidine-dependent growth was assessed. The GGAs do not bind to VAMP4 2–60 , while specific GGA interaction with CI-MPR was detected.

    Article Snippet: Since VAMP4 contains an acidic dileucine motif similar to CI-MPR , we tested for interaction between VAMP4 and the VHS domains of the GGAs.

    Techniques: Binding Assay, Incubation, Transformation Assay