2. Academic Validation
  3. 2.3 Å resolution cryo-EM structure of human p97 and mechanism of allosteric inhibition

2.3 Å resolution cryo-EM structure of human p97 and mechanism of allosteric inhibition

  • Science. 2016 Feb 19;351(6275):871-5. doi: 10.1126/science.aad7974.
Soojay Banerjee 1,  Alberto Bartesaghi 1,  Alan Merk 1,  Prashant Rao 1,  Stacie L Bulfer 2,  Yongzhao Yan 3,  Neal Green 4,  Barbara Mroczkowski 5,  R Jeffrey Neitz 2,  Peter Wipf 3,  Veronica Falconieri 1,  Raymond J Deshaies 6,  Jacqueline L S Milne 1,  Donna Huryn 3,  Michelle Arkin 2,  Sriram Subramaniam 7
Affiliations

Affiliations

  • 1 Laboratory of Cell Biology, National Cancer Institute, Bethesda, MD 20892, USA.
  • 2 Small Molecule Discovery Center, Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, CA 94143, USA.
  • 3 University of Pittsburgh Chemical Diversity Center, University of Pittsburgh, Pittsburgh, PA 15260, USA.
  • 4 Leidos Biomedical Research Inc., Frederick, MD 21702, USA.
  • 5 Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, MD 20892, USA.
  • 6 Division of Biology and Biological Engineering and Howard Hughes Medical Institute, California Institute of Technology, Pasadena, CA 91107, USA.
  • 7 Laboratory of Cell Biology, National Cancer Institute, Bethesda, MD 20892, USA. [email protected].
PMID: 26822609 DOI: 10.1126/science.aad7974
Abstract

p97 is a hexameric AAA+ adenosine triphosphatase (ATPase) that is an attractive target for Cancer drug development. We report cryo-electron microscopy (cryo-EM) structures for adenosine diphosphate (ADP)-bound, full-length, hexameric wild-type p97 in the presence and absence of an allosteric inhibitor at resolutions of 2.3 and 2.4 angstroms, respectively. We also report cryo-EM structures (at resolutions of ~3.3, 3.2, and 3.3 angstroms, respectively) for three distinct, coexisting functional states of p97 with occupancies of zero, one, or two molecules of adenosine 5'-O-(3-thiotriphosphate) (ATPγS) per protomer. A large corkscrew-like change in molecular architecture, coupled with upward displacement of the N-terminal domain, is observed only when ATPγS is bound to both the D1 and D2 domains of the protomer. These cryo-EM structures establish the sequence of nucleotide-driven structural changes in p97 at atomic resolution. They also enable elucidation of the binding mode of an allosteric small-molecule inhibitor to p97 and illustrate how inhibitor binding at the interface between the D1 and D2 domains prevents propagation of the conformational changes necessary for p97 function.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-123636
    99.85%, Allosteric p97 Inhibitor
    p97