5G4F

Structure of the ADP-bound VAT complex

Summary for 5G4F

• Entry DOI: 10.2210/pdb5g4f/pdb
• Related: 5G4G
• EMDB information: 3436
• Descriptor: VCP-LIKE ATPASE (1 entity in total)
• Functional Keywords: hydrolase, vat, proteasome, protein dynamics, unfoldase, conformations, aaa atpase
• Biological source: THERMOPLASMA ACIDOPHILUM
• Total number of polymer chains: 6
• Total formula weight: 486006.94

Authors

Huang, R.,Ripstein, Z.A.,Augustyniak, R.,Lazniewski, M.,Ginalski, K.,Kay, L.E.,Rubinstein, J.L. (deposition date: 2016-05-12, release date: 2016-07-27, Last modification date: 2024-05-08)

Primary citation

Huang, R.,Ripstein, Z.A.,Augustyniak, R.,Lazniewski, M.,Ginalski, K.,Kay, L.E.,Rubinstein, J.L.
Unfolding the Mechanism of the Aaa+ Unfoldase Vat by a Combined Cryo-Em, Solution NMR Study.
Proc.Natl.Acad.Sci.USA, 113:E4190-, 2016

PubMed Abstract: The AAA+ (ATPases associated with a variety of cellular activities) enzymes play critical roles in a variety of homeostatic processes in all kingdoms of life. Valosin-containing protein-like ATPase of Thermoplasma acidophilum (VAT), the archaeal homolog of the ubiquitous AAA+ protein Cdc48/p97, functions in concert with the 20S proteasome by unfolding substrates and passing them on for degradation. Here, we present electron cryomicroscopy (cryo-EM) maps showing that VAT undergoes large conformational rearrangements during its ATP hydrolysis cycle that differ dramatically from the conformational states observed for Cdc48/p97. We validate key features of the model with biochemical and solution methyl-transverse relaxation optimized spectroscopY (TROSY) NMR experiments and suggest a mechanism for coupling the energy of nucleotide hydrolysis to substrate unfolding. These findings illustrate the unique complementarity between cryo-EM and solution NMR for studies of molecular machines, showing that the structural properties of VAT, as well as the population distributions of conformers, are similar in the frozen specimens used for cryo-EM and in the solution phase where NMR spectra are recorded.

PubMed: 27402735
DOI: 10.1073/PNAS.1603980113

Experimental method

ELECTRON MICROSCOPY (7 Å)