Structural and functional characterization of the two phosphoinositide binding sites of PROPPINs, a β-propeller protein family
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- Roswitha Krick
- Department of Biochemistry II, Georg August University, D-37073 Göttingen, Germany;
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- Ricarda A. Busse
- Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, D-37077 Göttingen, Germany; and
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- Andreea Scacioc
- Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, D-37077 Göttingen, Germany; and
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- Milena Stephan
- Institute of Physical Chemistry, Georg August University, D-37077 Göttingen, Germany
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- Andreas Janshoff
- Institute of Physical Chemistry, Georg August University, D-37077 Göttingen, Germany
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- Michael Thumm
- Department of Biochemistry II, Georg August University, D-37073 Göttingen, Germany;
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- Karin Kühnel
- Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, D-37077 Göttingen, Germany; and
Abstract
<jats:p> β-propellers that bind polyphosphoinositides (PROPPINs), a eukaryotic WD-40 motif-containing protein family, bind via their predicted β-propeller fold the polyphosphoinositides PtdIns3P and PtdIns(3,5)P <jats:sub>2</jats:sub> using a conserved FRRG motif. PROPPINs play a key role in macroautophagy in addition to other functions. We present the 3.0-Å crystal structure of <jats:italic>Kluyveromyces lactis</jats:italic> Hsv2, which shares significant sequence homologies with its three <jats:italic>Saccharomyces cerevisiae</jats:italic> homologs Atg18, Atg21, and Hsv2. It adopts a seven-bladed β-propeller fold with a rare nonvelcro propeller closure. Remarkably, in the crystal structure, the two arginines of the FRRG motif are part of two distinct basic pockets formed by a set of highly conserved residues. In comprehensive in vivo and in vitro studies of ScAtg18 and ScHsv2, we define within the two pockets a set of conserved residues essential for normal membrane association, phosphoinositide binding, and biological activities. Our experiments show that PROPPINs contain two individual phosphoinositide binding sites. Based on docking studies, we propose a model for phosphoinositide binding of PROPPINs. </jats:p>
Journal
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- Proceedings of the National Academy of Sciences
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Proceedings of the National Academy of Sciences 109 (30), E2042-, 2012-07-02
Proceedings of the National Academy of Sciences
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Details 詳細情報について
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- CRID
- 1361418519154024064
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- ISSN
- 10916490
- 00278424
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- Data Source
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- Crossref