title: Nucleotide-dependent conformational changes of the AAA+ ATPase p97 revisited.
abstract: The ubiquitous AAA-ATPase p97 segregates ubiquitylated proteins from their molecular environment. Previous studies of the nucleotide-dependent conformational changes of p97 were inconclusive. Here, we determined its structure in the presence of ADP, AMP-PNP, or ATP-γS at 6.1-7.4 Å resolution using single particle cryo-electron microscopy. Both AAA domains, D1 and D2, assemble into essentially six-fold symmetrical rings. The pore of the D1-ring remains essentially closed under all nucleotide conditions, whereas the D2-ring shows an iris-like opening for ADP. The largest conformational changes of p97 are 'swinging motions' of the N-terminal domains, which may enable segregation of ubiquitylated substrates from their environment.
authors: Department of Molecular Structural Biology, Max-Planck Institute of Biochemistry, Martinsried, Germany.
authors: Jan M
authors: JM
authors: Schuller
authors: Department of Molecular Structural Biology, Max-Planck Institute of Biochemistry, Martinsried, Germany.
authors: Florian
authors: F
authors: Beck
authors: Biomolecular Mass Spectrometry and Proteomics and Netherlands Proteomics Center, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, The Netherlands.
authors: Philip
authors: P
authors: Lössl
authors: Biomolecular Mass Spectrometry and Proteomics and Netherlands Proteomics Center, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, The Netherlands.
authors: Albert J R
authors: AJ
authors: Heck
authors: Department of Molecular Structural Biology, Max-Planck Institute of Biochemistry, Martinsried, Germany.
authors: Friedrich
authors: http://orcid.org/0000-0002-6044-2746
authors: F
authors: Förster
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D008958:Models, Molecular; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism; D009711:Nucleotides / Q000494:pharmacology*; D017434:Protein Structure, Tertiary / Q000187:drug effects
chemical_list: D009687:Nuclear Proteins; D009711:Nucleotides; D000251:Adenosine Triphosphatases; C404942:p97 ATPase
keywords: AAA-ATPase; Cdc48; Cryo-EM; ERAD; protein quality control