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Structural analysis of the adaptor protein ClpS in complex with the N-terminal domain of ClpA

Nature Structural Biology volume 9, pages 906–911 (2002)Cite this article

Abstract

In Escherichia coli, protein degradation is performed by several proteolytic machines, including ClpAP. Generally, the substrate specificity of these machines is determined by chaperone components, such as ClpA. In some cases, however, the specificity is modified by adaptor proteins, such as ClpS. Here we report the 2.5 Å resolution crystal structure of ClpS in complex with the N-terminal domain of ClpA. Using mutagenesis, we demonstrate that two contact residues (Glu79 and Lys 84) are essential not only for ClpAS complex formation but also for ClpAPS-mediated substrate degradation. The corresponding residues are absent in the chaperone ClpB, providing a structural rationale for the unique specificity shown by ClpS despite the high overall similarity between ClpA and ClpB. To determine the location of ClpS within the ClpA hexamer, we modeled the N-terminal domain of ClpA onto a structurally defined, homologous AAA+ protein. From this model, we proposed a molecular mechanism to explain the ClpS-mediated switch in ClpA substrate specificity.

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Figure 1: Structure of ClpS in complex with the ClpA161
Figure 2: Structure of the N-domain of ClpA (ClpA161)
Figure 3: The formation of the ClpAS complex is essential for all ClpAS-mediated activities.
Figure 4: ClpS and the SsrA-tag do not compete directly for binding to ClpA161.

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Acknowledgements

We thank K. Truscott for critical reading of the manuscript.

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Author notes

  1. Bernd Bukau & David A. Dougan

    Present address: ZMBH, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg, D-69120, Germany

Authors and Affiliations

  1. Abteilung Membranbiochemie, MPI für Biochemie, Am Klopferspitz 18a, Martinsried, D-82152, Germany

    Kornelius Zeth

  2. EMBL outstation Grenoble, 6, rue Jules Horowitz, BP181, Grenoble, 38042, Cedex 9, France

    Kornelius Zeth, Raimond B. Ravelli & Stephen Cusack

  3. Institut für Biochemie, Universität Freiburg, Hermann-Herder-Strasse 7, Freiburg, D-79104, Germany

    Klaus Paal, Bernd Bukau & David A. Dougan

Authors
  1. Kornelius Zeth
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Correspondence to Kornelius Zeth or David A. Dougan.

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Zeth, K., Ravelli, R., Paal, K. et al. Structural analysis of the adaptor protein ClpS in complex with the N-terminal domain of ClpA. Nat Struct Mol Biol 9, 906–911 (2002). https://doi.org/10.1038/nsb869

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