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Dynamic structure of membrane-anchored Arf•GTP

Nature Structural & Molecular Biology volume 17, pages 876–881 (2010)Cite this article

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Abstract

ADP ribosylation factors (Arfs) are N-myristoylated GTP/GDP switch proteins that have key regulatory roles in vesicle transport in eukaryotic cells. ARFs execute their roles by anchoring to membrane surfaces, where they interact with other proteins to initiate budding and maturation of transport vesicles. However, existing structures of Arf•GTP are limited to nonmyristoylated and truncated forms with impaired membrane binding. We report a high-resolution NMR structure for full-length myristoylated yeast (Saccharomyces cerevisiae) Arf1 in complex with a membrane mimic. The two-domain structure, in which the myristoylated N-terminal helix is separated from the C-terminal domain by a flexible linker, suggests a level of adaptability in binding modes for the myriad of proteins with which Arf interacts and allows predictions of specific lipid binding sites on some of these proteins.

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Figure 1: Structure of myr-yArf1•GTP in a bicelle solution.
Figure 2: Lipid interaction of myr-yArf1 studied through rotational correction times (τc).
Figure 3: Ensemble structural fitting to RDCs and PREs.
Figure 4: Reweighted atomic density maps of the C-terminal domain (Glu17–Leu177) showing the space on the membrane surface that is sampled by the three dynamic states.
Figure 5: Modeling of Arf complexes on the membrane.

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Acknowledgements

The authors thank M. Demarco (Complex Carbohydrate Research Center, Univ. of Georgia) for kindly providing the model membrane structure and C. Schwieters (Center for Information Technology, US National Institutes of Health) for kindly providing the prerelease version of Xplor-NIH that supports ambiguous RDC assignment. This work was supported by a grant from the US National Institutes of Health (GM61268).

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Authors and Affiliations

  1. Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia, USA

    Yizhou Liu & James H Prestegard

  2. Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA

    Richard A Kahn

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Y.L. produced the samples and collected and analyzed all data; J.H.P. and R.A.K. devised the project and jointly contributed interpretation of data and drafting of the manuscript.

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Correspondence to Richard A Kahn or James H Prestegard.

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Liu, Y., Kahn, R. & Prestegard, J. Dynamic structure of membrane-anchored Arf•GTP. Nat Struct Mol Biol 17, 876–881 (2010). https://doi.org/10.1038/nsmb.1853

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