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The calcium binding protein ALG-2 binds and stabilizes Scotin, a p53-inducible gene product localized at the endoplasmic reticulum membrane.

Draeby I, Woods YL, la Cour JM, Mollerup J, Bourdon JC, Berchtold MW - Arch. Biochem. Biophys. (2007)

Bottom Line: In this study we identified Scotin as a novel ALG-2 target protein containing 6 PXY and 4 PYP repeats, earlier identified in the ALG-2 binding regions of AIP1/ALIX and TSG101, respectively.Overexpression of ALG-2 led to accumulation of Scotin in MCF7 and H1299 cells.In vitro and in vivo binding of ALG-2 to Scotin was demonstrated to be strictly calcium dependent indicating a role of this interaction in calcium signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, University of Copenhagen, Biocenter, Ole Maaløes Vej 5, 2200 Copenhagen N, Denmark.

ABSTRACT
ALG-2 (apoptosis linked gene 2 product) is a calcium binding protein for which no clear cellular function has been established. In this study we identified Scotin as a novel ALG-2 target protein containing 6 PXY and 4 PYP repeats, earlier identified in the ALG-2 binding regions of AIP1/ALIX and TSG101, respectively. An in vitro synthesized C-terminal fragment of Scotin bound specifically to immobilized recombinant ALG-2 and tagged ALG-2 and Scotin were shown by immunoprecipitation to interact in MCF7 and U2OS cell lines. Furthermore ALG-2 bound to endogenous Scotin in extracts from mouse NIH3T3 cells. Overexpression of ALG-2 led to accumulation of Scotin in MCF7 and H1299 cells. In vitro and in vivo binding of ALG-2 to Scotin was demonstrated to be strictly calcium dependent indicating a role of this interaction in calcium signaling pathways.

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(a) Structure of Scotin and (b) sequence alignment by the ClustalW program of the proline/tyrosine-rich regions of AIP1/Alix (Accession No. NP_037506, TSG101, Accession No. NP_006283 and Scotin (Accession No. NP_057563). Highlighted residues were either identical in the three sequences or in two sequences.
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fig1: (a) Structure of Scotin and (b) sequence alignment by the ClustalW program of the proline/tyrosine-rich regions of AIP1/Alix (Accession No. NP_037506, TSG101, Accession No. NP_006283 and Scotin (Accession No. NP_057563). Highlighted residues were either identical in the three sequences or in two sequences.

Mentions: Elucidation of the binding of ALG-2 to TSG101, AIP1/Alix, Annexin 7 and 11 (reviewed in [3]) encouraged database searches for protein sequences containing similar proline/tyrosine-rich regions with the goal to find novel ALG-2 binding partners. One of the candidate proteins found by this in silico approach was Scotin (Fig. 1a). Comparisons of the proline/tyrosine-rich region of Scotin with known ALG-2 interaction partners revealed that the proline-rich region of Scotin does contain six PXY repeats (residues 166–168, 171–173, 196–198, 206–208, 225–227, 229–231) as described for the ALG-2 binding region in AIP1/ALIX [9] and 4 PYP repeats (residues 157–159, 189–191, 197–199, 219–221) as well as 1 YPP sequence (residues 194–196), as described for the ALG-2 binding region in TSG101 [8] (Fig. 1b).


The calcium binding protein ALG-2 binds and stabilizes Scotin, a p53-inducible gene product localized at the endoplasmic reticulum membrane.

Draeby I, Woods YL, la Cour JM, Mollerup J, Bourdon JC, Berchtold MW - Arch. Biochem. Biophys. (2007)

(a) Structure of Scotin and (b) sequence alignment by the ClustalW program of the proline/tyrosine-rich regions of AIP1/Alix (Accession No. NP_037506, TSG101, Accession No. NP_006283 and Scotin (Accession No. NP_057563). Highlighted residues were either identical in the three sequences or in two sequences.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2691584&req=5

fig1: (a) Structure of Scotin and (b) sequence alignment by the ClustalW program of the proline/tyrosine-rich regions of AIP1/Alix (Accession No. NP_037506, TSG101, Accession No. NP_006283 and Scotin (Accession No. NP_057563). Highlighted residues were either identical in the three sequences or in two sequences.
Mentions: Elucidation of the binding of ALG-2 to TSG101, AIP1/Alix, Annexin 7 and 11 (reviewed in [3]) encouraged database searches for protein sequences containing similar proline/tyrosine-rich regions with the goal to find novel ALG-2 binding partners. One of the candidate proteins found by this in silico approach was Scotin (Fig. 1a). Comparisons of the proline/tyrosine-rich region of Scotin with known ALG-2 interaction partners revealed that the proline-rich region of Scotin does contain six PXY repeats (residues 166–168, 171–173, 196–198, 206–208, 225–227, 229–231) as described for the ALG-2 binding region in AIP1/ALIX [9] and 4 PYP repeats (residues 157–159, 189–191, 197–199, 219–221) as well as 1 YPP sequence (residues 194–196), as described for the ALG-2 binding region in TSG101 [8] (Fig. 1b).

Bottom Line: In this study we identified Scotin as a novel ALG-2 target protein containing 6 PXY and 4 PYP repeats, earlier identified in the ALG-2 binding regions of AIP1/ALIX and TSG101, respectively.Overexpression of ALG-2 led to accumulation of Scotin in MCF7 and H1299 cells.In vitro and in vivo binding of ALG-2 to Scotin was demonstrated to be strictly calcium dependent indicating a role of this interaction in calcium signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, University of Copenhagen, Biocenter, Ole Maaløes Vej 5, 2200 Copenhagen N, Denmark.

ABSTRACT
ALG-2 (apoptosis linked gene 2 product) is a calcium binding protein for which no clear cellular function has been established. In this study we identified Scotin as a novel ALG-2 target protein containing 6 PXY and 4 PYP repeats, earlier identified in the ALG-2 binding regions of AIP1/ALIX and TSG101, respectively. An in vitro synthesized C-terminal fragment of Scotin bound specifically to immobilized recombinant ALG-2 and tagged ALG-2 and Scotin were shown by immunoprecipitation to interact in MCF7 and U2OS cell lines. Furthermore ALG-2 bound to endogenous Scotin in extracts from mouse NIH3T3 cells. Overexpression of ALG-2 led to accumulation of Scotin in MCF7 and H1299 cells. In vitro and in vivo binding of ALG-2 to Scotin was demonstrated to be strictly calcium dependent indicating a role of this interaction in calcium signaling pathways.

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