Scientific article

Recognition, targeting, and hydrolysis of the lambda O replication protein by the ClpP/ClpX protease

Published inThe Journal of biological chemistry, vol. 274, no. 20, p. 13999-14005
Publication date1999

It has previously been established that sequences at the C termini of polypeptide substrates are critical for efficient hydrolysis by the ClpP/ClpX ATP-dependent protease. We report for the bacteriophage lambda O replication protein, however, that N-terminal sequences play the most critical role in facilitating proteolysis by ClpP/ClpX. The N-terminal portion of lambda O is degraded at a rate comparable with that of wild type O protein, whereas the C-terminal domain of O is hydrolyzed at least 10-fold more slowly. Consistent with these results, deletion of the first 18 amino acids of lambda O blocks degradation of the N-terminal domain, whereas proteolysis of the O C-terminal domain is only slightly diminished as a result of deletion of the C-terminal 15 amino acids. We demonstrate that ClpX retains its capacity to bind to the N-terminal domain following removal of the first 18 amino acids of O. However, ClpX cannot efficiently promote the ATP-dependent binding of this truncated O polypeptide to ClpP, the catalytic subunit of the ClpP/ClpX protease. Based on our results with lambda O protein, we suggest that two distinct structural elements may be required in substrate polypeptides to enable efficient hydrolysis by the ClpP/ClpX protease: (i) a ClpX-binding site, which may be located remotely from substrate termini, and (ii) a proper N- or C-terminal sequence, whose exposure on the substrate surface may be induced by the binding of ClpX.

  • Adenosine Triphosphatases/*metabolism
  • Amino Acid Sequence
  • Bacteriophage lambda/metabolism/*physiology
  • Binding Sites
  • Endopeptidase Clp
  • Enzyme-Linked Immunosorbent Assay
  • Hydrolysis
  • Kinetics
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Serine Endopeptidases/*metabolism
  • Viral Proteins/genetics/*metabolism
  • *Virus Replication
Citation (ISO format)
GONCIARZ-SWIATEK, M. et al. Recognition, targeting, and hydrolysis of the lambda O replication protein by the ClpP/ClpX protease. In: The Journal of biological chemistry, 1999, vol. 274, n° 20, p. 13999–14005.
Updates (1)
ISSN of the journal0021-9258

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