Structural basis for distinct operational modes and protease activation in AAA+ protease Lon

Sci Adv. 2020 May 20;6(21):eaba8404. doi: 10.1126/sciadv.aba8404. eCollection 2020 May.


Substrate-bound structures of AAA+ protein translocases reveal a conserved asymmetric spiral staircase architecture wherein a sequential ATP hydrolysis cycle drives hand-over-hand substrate translocation. However, this configuration is unlikely to represent the full conformational landscape of these enzymes, as biochemical studies suggest distinct conformational states depending on the presence or absence of substrate. Here, we used cryo-electron microscopy to determine structures of the Yersinia pestis Lon AAA+ protease in the absence and presence of substrate, uncovering the mechanistic basis for two distinct operational modes. In the absence of substrate, Lon adopts a left-handed, "open" spiral organization with autoinhibited proteolytic active sites. Upon the addition of substrate, Lon undergoes a reorganization to assemble an enzymatically active, right-handed "closed" conformer with active protease sites. These findings define the mechanistic principles underlying the operational plasticity required for processing diverse protein substrates.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATPases Associated with Diverse Cellular Activities / metabolism
  • Adenosine Triphosphate / metabolism
  • Catalytic Domain
  • Cryoelectron Microscopy
  • Endopeptidases*
  • Peptide Hydrolases* / metabolism
  • Proteolysis


  • Adenosine Triphosphate
  • Endopeptidases
  • Peptide Hydrolases
  • ATPases Associated with Diverse Cellular Activities