Abstract
P97 protein, also referred to as valosin-containing protein (VCP), is an AAA-ATPase (ATPase associated with a variety of cellular activities) that mediates vital cellular activities with the cooperation of many cofactors. A group of cofactors interact with the N-terminal domain of P97 (P97N) through their Arg/Lys-rich peptide motifs. We investigated the interactions between P97 and these motifs, including VCP-binding motif (VBM) and VCP-interacting motif (VIM). The solution structures of the VBM motif from HRD1 and the VIM motif from SVIP are both comprised mainly of a single α-helix. The VIM motifs generally have stronger P97N-binding affinities than the VBMs, and SVIP (VIM) can compete with HRD1-VBM for the interaction, providing a possibility that VIM-containing proteins (such as SVIP) act as competitors against VBM-containing proteins (such as HRD1) for interacting with P97. Based on biochemical features of the VBM motifs, we also identified NUB1L (NEDD8 ultimate buster-1 long) as a novel VBM-containing protein, which is involved in proteasomal degradation of NEDD8 through the P97 pathway.
Keywords:
AAA; ATPase associated with a variety of cellular activities; Arginine/lysine-rich; BS1; ERAD; ITC; Interaction; NMR; P97-ND1; P97/VCP; P97N; Structure; VBM; VCP; VCP-binding motif; VCP-interacting motif; VIM; binding site 1; endoplasmic reticulum-associated degradation; isothermal titration calorimetry; nuclear magnetic resonance; the N and D1 domains of P97 (residues 1–458); the N-terminal domain of P97 (residues 1–213); valosin-containing protein.
© 2013.
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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Adenosine Triphosphatases / chemistry
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Adenosine Triphosphatases / genetics
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Adenosine Triphosphatases / metabolism*
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Amino Acid Motifs
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Arginine / chemistry
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Arginine / genetics
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Arginine / metabolism
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Carrier Proteins / chemistry
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Cell Cycle Proteins / chemistry
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Cell Cycle Proteins / genetics
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Cell Cycle Proteins / metabolism*
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HEK293 Cells
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Humans
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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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NEDD8 Protein
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Peptides / chemistry
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Peptides / genetics
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Peptides / metabolism*
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Phosphate-Binding Proteins
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Proline / chemistry
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Proline / genetics
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Proline / metabolism
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Proteasome Endopeptidase Complex / chemistry
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Proteasome Endopeptidase Complex / genetics
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Proteasome Endopeptidase Complex / metabolism
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Protein Binding
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Proteolysis
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Transcription Factors / chemistry
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Transcription Factors / genetics
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Transcription Factors / metabolism*
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Ubiquitin-Protein Ligases / chemistry
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Ubiquitin-Protein Ligases / genetics
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Ubiquitin-Protein Ligases / metabolism*
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Ubiquitins / chemistry
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Ubiquitins / genetics
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Ubiquitins / metabolism
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Valosin Containing Protein
Substances
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Adaptor Proteins, Signal Transducing
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Carrier Proteins
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Cell Cycle Proteins
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Membrane Proteins
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NEDD8 Protein
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NEDD8 protein, human
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NUB1 protein, human
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Peptides
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Phosphate-Binding Proteins
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SVIP protein, human
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Transcription Factors
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Ubiquitins
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Arginine
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Proline
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SYVN1 protein, human
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Ubiquitin-Protein Ligases
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Proteasome Endopeptidase Complex
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Adenosine Triphosphatases
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VCP protein, human
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Valosin Containing Protein