Identification and structural characterization of two 14-3-3 binding sites in the human peptidylarginine deiminase type VI.
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.
Chemical Genomics Centre of the Max-Planck-Society, Germany.
Rolf
R
Rose
Micheline
M
Rose
Christian
C
Ottmann
D048948:14-3-3 Proteins / Q000737:chemistry*; D000595:Amino Acid Sequence; D001665:Binding Sites; D014408:Biomarkers, Tumor / Q000737:chemistry*; D018360:Crystallography, X-Ray; D005092:Exonucleases / Q000737:chemistry*; D005095:Exoribonucleases; D006801:Humans; D006867:Hydrolases / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry; D010766:Phosphorylation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D000080065:Protein-Arginine Deiminase Type 6; D000076342:Protein-Arginine Deiminases; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000737:chemistry
D048948:14-3-3 Proteins; D014408:Biomarkers, Tumor; D010446:Peptide Fragments; D010750:Phosphoproteins; D038744:Ribosomal Protein S6 Kinases, 90-kDa; D006867:Hydrolases; D005092:Exonucleases; D005095:Exoribonucleases; C413442:SFN protein, human; C488263:PADI6 protein, human; D000080065:Protein-Arginine Deiminase Type 6; D000076342:Protein-Arginine Deiminases