Abstract
The POTE gene family is composed of 13 paralogs and likely evolved by duplications and remodeling of the human genome. One common property of POTE proteins is their localization on the inner aspect of the plasma membrane. To determine the structural elements required for membrane localization, we expressed mutants of different POTEs in 293T cells as EGFP fusion proteins. We also tested their palmitoylation by a biotin-switch assay. Our data indicate that the membrane localizations of different POTEs are mediated by similar 3-4 short cysteine rich repeats (CRRs) near the amino-terminuses and that palmitoylation on paired cysteine residues in each CRR motif is responsible for the localization. Multiple palmitoylation in the small CRRs can result in the strong association of whole POTEs with plasma membrane.
Publication types
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Research Support, N.I.H., Intramural
MeSH terms
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Amino Acid Motifs
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Amino Acid Sequence
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Cell Line
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Cell Membrane / metabolism*
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Cysteine / chemistry
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Cysteine / genetics
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Cysteine / metabolism
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Humans
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Immunoblotting
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Lipoylation / drug effects
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Microscopy, Confocal
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Mutagenesis
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Mutation
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Palmitates / pharmacology
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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Protein Structure, Secondary
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Protein Transport / drug effects
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Protein Transport / genetics
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism*
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Transfection
Substances
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Membrane Proteins
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Palmitates
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Protein Isoforms
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Recombinant Fusion Proteins
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enhanced green fluorescent protein
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Green Fluorescent Proteins
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2-bromopalmitate
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Cysteine