Characterization of the N-terminal repeat domain of Escherichia coli ClpA-A class I Clp/HSP100 ATPase

Protein Sci. 2001 Mar;10(3):551-9. doi: 10.1110/ps.41401.

Abstract

The ClpA, ClpB, and ClpC subfamilies of the Clp/HSP100 ATPases contain a conserved N-terminal region of approximately 150 residues that consists of two approximate sequence repeats. This sequence from the Escherichia coli ClpA enzyme is shown to encode an independent structural domain (the R domain) that is monomeric and approximately 40% alpha-helical. A ClpA fragment lacking the R domain showed ATP-dependent oligomerization, protein-stimulated ATPase activity, and the ability to complex with the ClpP peptidase and mediate degradation of peptide and protein substrates, including casein and ssrA-tagged proteins. Compared with the activities of the wild-type ClpA, however, those of the ClpA fragment missing the R domain were reduced. These results indicate that the R domain is not required for the basic recognition, unfolding, and translocation functions that allow ClpA-ClpP to degrade some protein substrates, but they suggest that it may play a role in modulating these activities.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphatases / chemistry*
  • Adenosine Triphosphatases / metabolism*
  • Amino Acid Sequence
  • Biophysical Phenomena
  • Biophysics
  • Endopeptidase Clp
  • Enzyme Activation
  • Escherichia coli / classification
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli Proteins*
  • Hydrolysis
  • Mutagenesis / genetics*
  • Protein Structure, Tertiary
  • Sequence Deletion / genetics
  • Sequence Homology
  • Serine Endopeptidases / chemistry*
  • Serine Endopeptidases / metabolism*
  • Terminal Repeat Sequences*

Substances

  • Escherichia coli Proteins
  • Serine Endopeptidases
  • ClpA protease, E coli
  • Endopeptidase Clp
  • Adenosine Triphosphatases