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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Animals
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Apoptosis Regulatory Proteins / metabolism*
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Calcium-Binding Proteins / metabolism*
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Carrier Proteins
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DNA-Binding Proteins / metabolism
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Endoplasmic Reticulum / metabolism*
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Endosomal Sorting Complexes Required for Transport
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Guanylate Kinases
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Humans
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Intracellular Membranes / metabolism*
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Membrane Proteins / metabolism*
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Mice
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NIH 3T3 Cells
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Nuclear Proteins / metabolism*
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Proteins / metabolism
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Recombinant Fusion Proteins / chemistry
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Transcription Factors / metabolism*
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Tumor Suppressor Protein p53 / metabolism*
Substances
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Adaptor Proteins, Signal Transducing
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Apoptosis Regulatory Proteins
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Calcium-Binding Proteins
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Carrier Proteins
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DNA-Binding Proteins
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Endosomal Sorting Complexes Required for Transport
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Membrane Proteins
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Nuclear Proteins
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PDCD6 protein, human
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Pdcd6 protein, mouse
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Proteins
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Recombinant Fusion Proteins
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SHISA5 protein, human
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Scotin protein, mouse
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TP53 protein, human
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Transcription Factors
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Tsg101 protein
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Tumor Suppressor Protein p53
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Guanylate Kinases
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MAGI2 protein, human