Abstract
Appropriate targeting of matrix proteins to peroxisomes is mainly directed by two types of peroxisomal targeting signals, PTS1 and PTS2. Although the basis of PTS1 recognition has been revealed by structural studies, that of PTS2 recognition remains elusive. Here we present the crystal structure of a heterotrimeric PTS2-recognition complex from Saccharomyces cerevisiae, containing Pex7p, the C-terminal region of Pex21p and the PTS2 of the peroxisomal 3-ketoacyl-CoA thiolase. Pex7p forms a β-propeller structure and provides a platform for cooperative interactions with both the amphipathic PTS2 helix and Pex21p. The C-terminal region of Pex21p directly covers the hydrophobic surfaces of both Pex7p and PTS2, and the resulting hydrophobic core is the primary determinant of stable complex formation. Together with in vivo and in vitro functional assays of Pex7p and Pex21p variants, our findings reveal the molecular mechanism of PTS2 recognition.
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Acknowledgements
We thank H. Hirai for helpful advice on plasmid construction, protein purification and crystallization. We thank Y. Matsuura for critical reading of the manuscript. This work was supported by the Targeted Proteins Research Program of the Ministry of Education, Culture, Sports, Science, and Technology, Japan (H.K.). The synchrotron radiation experiments were performed at SPring-8 BL41XU (2011A1210, 2011A1289).
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D.P. generated plasmids, proteins and crystals. D.P. and T.N. collected diffraction data and solved the structure. D.P. performed pulldown experiments and in vivo functional assays. H.K. supervised the project. D.P. and H.K. wrote the paper.
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Pan, D., Nakatsu, T. & Kato, H. Crystal structure of peroxisomal targeting signal-2 bound to its receptor complex Pex7p–Pex21p. Nat Struct Mol Biol 20, 987–993 (2013). https://doi.org/10.1038/nsmb.2618
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DOI: https://doi.org/10.1038/nsmb.2618
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