Escherichia coli ClpB is a non-processive polypeptide translocase

Academic Article

Abstract

  • © 2015 Authors; published by Portland Press Limited. Escherichia coli caseinolytic protease (Clp)B is a hexameric AAA + [expanded superfamily of AAA (ATPase associated with various cellular activities)] enzyme that has the unique ability to catalyse protein disaggregation. Such enzymes are essential for proteome maintenance. Based on structural comparisons to homologous enzymes involved in ATP-dependent proteolysis and clever protein engineering strategies, it has been reported that ClpB translocates polypeptide through its axial channel. Using single-turnover fluorescence and anisotropy experiments we show that ClpB is a non-processive polypeptide translocase that catalyses disaggregation by taking one or two translocation steps followed by rapid dissociation. Using single-turnover FRET experiments we show that ClpB containing the IGL loop from ClpA does not translocate substrate through its axial channel and into ClpP for proteolytic degradation. Rather, ClpB containing the IGL loop dysregulates ClpP leading to non-specific proteolysis reminiscent of ADEP (acyldepsipeptide) dysregulation. Our results support a molecular mechanism where ClpB catalyses protein disaggregation by tugging and releasing exposed tails or loops.
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    Digital Object Identifier (doi)

    Author List

  • Li T; Weaver CL; Lin J; Duran EC; Miller JM; Lucius AL
  • Start Page

  • 39
  • End Page

  • 52
  • Volume

  • 470
  • Issue

  • 1