PDBsum entry 5g4g

Hydrolase

PDB id: 5g4g

Contents

Protein chains

(+ 0 more) 721 a.a.

PDB id:

5g4g

Name:

Hydrolase

Title:

Structure of the atpgs-bound vat complex

Structure:

Vcp-like atpase. Chain: a, b, c, d, e, f. Engineered: yes

Source:

Thermoplasma acidophilum. Organism_taxid: 2303. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Authors:

R.Huang,Z.A.Ripstein,R.Augustyniak,M.Lazniewski,K.Ginalski,L.E.Kay, J.L.Rubinstein

Key ref:

R.Huang et al. (2016). Unfolding the mechanism of the AAA+ unfoldase VAT by a combined cryo-EM, solution NMR study. Proc Natl Acad Sci U S A, 113, E4190. PubMed id: 27402735 DOI: 10.1073/pnas.1603980113

Date:

12-May-16 Release date: 27-Jul-16

Protein chains

O05209  (VAT_THEAC) -  VCP-like ATPase from Thermoplasma acidophilum (strain ATCC 25905 / DSM 1728 / JCM 9062 / NBRC 15155 / AMRC-C165)

Seq: 745 a.a.

Struc: 721 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1073/pnas.1603980113

Proc Natl Acad Sci U S A 113:E4190 (2016)

PubMed id: 27402735

Unfolding the mechanism of the AAA+ unfoldase VAT by a combined cryo-EM, solution NMR study.

R.Huang, Z.A.Ripstein, R.Augustyniak, M.Lazniewski, K.Ginalski, L.E.Kay, J.L.Rubinstein.

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.