Crystal structure of the Saccharomyces cerevisiae phosphatidylinositol- transfer protein

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Abstract

The yeast phosphatidylinositol-transfer protein (Sec14) catalyses exchange of phosphatidylinositol and phosphatidylcholine between membrane bilayers in vitro1,2. In vivo, Sec14 activity is essential for vesicle budding from the Golgi complex3. Here we report a three-dimensional structure for Sec14 at 2.5 Å resolution. Sec14 consists of twelve α-helices, six β-strands, eight 310-helices and has two distinct domains. The carboxy-terminal domain forms a hydrophobic pocket which, in the crystal ructure, is occupied by two molecules of n-octyl-β-D-glucopyranoside and represents the phospholipid-binding domain. This pocket is reinforced by a string motif whose disruption in a sec14 temperature-sensitive mutant results in destabilization of the phospholipid-binding domain. Finally, we have identified an unusual surface helix that may play a critical role in driving Sec14-mediated phospholipid exchange. From this structure, we derive the first molecular clues into how a phosphatidylinositol-transfer protein functions.

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Figure 1: Sec14 structure.
Figure 2: Bound detergent in the Sec14 structure.
Figure 3: Bulldozer model for Sec14-mediated phospholipid exchange.

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Acknowledgements

We thank J. Collawn for helpful discussions and critical reading of the manuscript and D. Malehorn for help with site-directed mutagenesis. We thank J. Tsao and Y. Luo for their help in data collection. This work was supported by a grant from the NIH to V.A.B. and NASA to M.L. The X-ray crystallographic coordinates have been deposited in the BNL Protein Data Bank (accession number 1AUA).

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Correspondence to Vytas A. Bankaitis or Ming Luo.

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