Abstract
The regulation and function of peptidylarginine deiminase isoform VI (PAD6), which is a highly abundant protein associated with the cytoplasmic lattices in mammalian oocytes, is poorly understood so far. It has been shown previously, that 14-3-3 proteins, a class of regulatory adapter proteins ubiquitous in eukaryotes, bind to PAD6 in vivo in a phosphorylation dependent manner. Here we identify possible 14-3-3 binding sites in human PAD6 by in silico methods, looking for conserved, surface exposed serine residues. Two of these sites were confirmed as 14-3-3 binding sites by fluorescence polarization competition and X-ray crystallography. We furthermore suggest a role of RSK-type kinases in the phosphorylation of one of these two binding sites and provide evidence in the form of in vitro kinase assays with p70S6 kinase and RSK1.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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14-3-3 Proteins / chemistry*
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Amino Acid Sequence
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Binding Sites
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Biomarkers, Tumor / chemistry*
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Crystallography, X-Ray
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Exonucleases / chemistry*
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Exoribonucleases
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Humans
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Hydrolases / chemistry*
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Hydrophobic and Hydrophilic Interactions
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Models, Molecular
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Molecular Sequence Data
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Peptide Fragments / chemistry
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Phosphoproteins / chemistry
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Phosphorylation
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Protein Binding
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Protein Interaction Domains and Motifs
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Protein Processing, Post-Translational
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Protein Structure, Secondary
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Protein-Arginine Deiminase Type 6
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Protein-Arginine Deiminases
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Ribosomal Protein S6 Kinases, 90-kDa / chemistry
Substances
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14-3-3 Proteins
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Biomarkers, Tumor
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Peptide Fragments
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Phosphoproteins
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Ribosomal Protein S6 Kinases, 90-kDa
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Hydrolases
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Exonucleases
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Exoribonucleases
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SFN protein, human
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PADI6 protein, human
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Protein-Arginine Deiminase Type 6
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Protein-Arginine Deiminases