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Structure of the Janus-faced C2B domain of rabphilin

Nature Cell Biology volume 1, pages 106–112 (1999)Cite this article

Abstract

C2 domains are widespread protein modules that often occur as tandem repeats in many membrane-trafficking proteins such as synaptotagmin and rabphilin. The first and second C2 domains (C2A and C2B, respectively) have a high degree of homology but also specific differences. The structure of the C2A domain of synaptotagmin I has been extensively studied but little is known about the C2B domains. We have used NMR spectroscopy to determine the solution structure of the C2B domain of rabphilin. The overall structure of the C2B domain is very similar to that of other C2 domains, with a rigid β-sandwich core and loops at the top (where Ca2+ binds) and the bottom. Surprisingly, a relatively long α-helix is inserted at the bottom of the domain and is conserved in all C2B domains. Our results, together with the Ca2+-independent interactions observed for C2B domains, indicate that these domains have a Janus-faced nature, with a Ca2+-binding top surface and a Ca2+-independent bottom surface.

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Figure 1: The C2B domain binds Ca2+ in the absence of phospholipids.
Figure 2: Ca2+ binding to the C2B domain monitored by tryptophan fluorescence.
Figure 3: Solution structure of the C2B domain.
Figure 4: Sequence alignment of the C-terminal regions of C2 domains from proteins containing two consecutive C2 domains.
Figure 6: Taxonomy of C2 domains.
Figure 7: Comparison of the Ca2+-binding mode of the C2B domain with those of other C2 domains.
Figure 5: Packing of helix 2 with the rest of the C2B domain.

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Acknowledgements

We thank L. Kay for providing pulse sequences for all triple-resonance experiments, and B. Sutton and A. Brunger for communicating results before publication. J.U. was a fellow from the Direccio General de Recerca, Generalitat de Catalunya, Spain, and J.G. was a fellow from the Departamento de Educacion, Universidades e Investigacion, Gobierno Vasco, Spain. This work was supported by NIH grant NS33731.

Correspondence and requests for materials should be addressed to J.R. The 20 structures of the rabphilin C2B domain have been deposited in the Protein Data Bank with accession number 3rpb.

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  1. M. Paige Nittler

    Present address: University of California at San Francisco, 513 Parnassus Avenue, Box 0450, San Francisco, California, 94143, USA

Authors and Affiliations

  1. Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard , Dallas, 75235, Texas, USA

    Josep Ubach, Jesús García, M. Paige Nittler & Josep Rizo

  2. Department of Molecular Genetics, Howard Hughes Medical Institute and Center for Basic Neuroscience, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas , 75235, Texas, USA

    Thomas C. Südhof

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Ubach, J., García, J., Nittler, M. et al. Structure of the Janus-faced C2B domain of rabphilin. Nat Cell Biol 1, 106–112 (1999). https://doi.org/10.1038/10076

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