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PAN, the proteasome-activating nucleotidase from archaebacteria, is a protein-unfolding molecular chaperone

Nature Cell Biology volume 2, pages 833–839 (2000)Cite this article

Abstract

The proteasome-activating nucleotidase (PAN) from Methanococcus jannaschii is a complex of relative molecular mass 650,000 that is homologous to the ATPases in the eukaryotic 26S proteasome. When mixed with 20S archaeal proteasomes and ATP, PAN stimulates protein degradation. Here we show that PAN reduces aggregation of denatured proteins and enhances their refolding. These processes do not require ATP hydrolysis, although ATP binding enhances the ability of PAN to prevent aggregation. PAN also catalyses the unfolding of the green fluorescent protein with an 11-residue ssrA extension at its carboxy terminus (GFP11). This unfolding requires ATP hydrolysis, and is linked to GFP11 degradation when 20S proteasomes are also present. This unfolding activity seems to be essential for ATP-dependent proteolysis, although PAN may function by itself as a molecular chaperone.

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Figure 1: PAN reduces protein aggregation.
Figure 2: PAN enhances refolding of denatured GDH.
Figure 3: The ATP- and AMP-PNP-bound forms of PAN are more effective in preventing protein aggregation.
Figure 4: Effect of addition of 20S proteasomes on the ability of PAN to prevent protein aggregation.
Figure 5: PAN catalyses ATP-dependent unfolding of GFP11.
Figure 6: PAN stimulates ATP-dependent degradation of GFP11 by 20S proteasomes.

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Acknowledgements

We thank A. L. Horwich for the GFP11 expression plasmid, P. Silver for native GFP expression plasmid, and A. Kisselev and A. Navon for the purified 20S proteasome. We also thank members of our laboratory for critical reading of the manuscript. This work was supported by research grants from the National Institutes of Health (to A.L.G.; GM46147 and GM51923) and by postdoctoral fellowships from Association pour la Recherche contre le Cancer and Human Frontier Science Program Organization (to N.B.).

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  1. Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Nadia Benaroudj & Alfred L. Goldberg

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  1. Nadia Benaroudj
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Correspondence to Alfred L. Goldberg.

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Benaroudj, N., Goldberg, A. PAN, the proteasome-activating nucleotidase from archaebacteria, is a protein-unfolding molecular chaperone. Nat Cell Biol 2, 833–839 (2000). https://doi.org/10.1038/35041081

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