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Visualization of an unstable coiled coil from the scallop myosin rod

Nature volume 424, pages 341–345 (2003)Cite this article

Abstract

α-Helical coiled coils in muscle exemplify simplicity and economy of protein design: small variations in sequence lead to remarkable diversity in cellular functions1,2. Myosin II is the key protein in muscle contraction, and the molecule's two-chain α-helical coiled-coil rod region—towards the carboxy terminus of the heavy chain—has unusual structural and dynamic features. The amino-terminal subfragment-2 (S2) domains of the rods can swing out from the thick filament backbone at a hinge in the coiled coil, allowing the two myosin ‘heads’ and their motor domains to interact with actin and generate tension3. Most of the S2 rod appears to be a flexible coiled coil, but studies suggest that the structure at the N-terminal region is unstable4,5,6, and unwinding or bending of the α-helices near the head–rod junction seems necessary for many of myosin's functional properties7,8. Here we show the physical basis of a particularly weak coiled-coil segment by determining the 2.5-Å-resolution crystal structure of a leucine-zipper-stabilized fragment of the scallop striated-muscle myosin rod adjacent to the head–rod junction. The N-terminal 14 residues are poorly ordered; the rest of the S2 segment forms a flexible coiled coil with poorly packed core residues. The unusual absence of interhelical salt bridges here exposes apolar core atoms to solvent.

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Figure 1: S2N51 is a flexible coiled coil.
Figure 2: Scallop S2 displays non-close-packed regions in the core and conformationally variable ‘a’-position lysine residues.
Figure 3: The absence of interhelical salt bridges between the ‘g’ and ‘e’ positions (purple) exposes core residues (green) to solvent.
Figure 4: The N-terminal region of molluscan S2 (sequences 1 and 2) displays in an exaggerated way the moderately low level of coiled-coil-stabilizing features present in S2 of various myosins.

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Acknowledgements

We thank D. A. D. Parry, A. G. Szent-Györgyi and H. E. Huxley for a critical reading of the manuscript, and the staff of the Cornell High Energy Synchrotron Source for assistance with data collection. This work has been supported by grants to C.C. from the National Institutes of Health and the Muscular Dystrophy Association, and to L.N. from the Hungarian Scientific Research Fund (OTKA).

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  1. Yu Li

    Present address: Boston Biomedical Research Institute, 64 Grove Street, Watertown, Massachusetts, 02472, USA

Authors and Affiliations

  1. Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts, 02454-9110, USA

    Yu Li, Jerry H. Brown, Ludmilla Reshetnikova & Carolyn Cohen

  2. Biophysics and Structural Biology Program, Brandeis University, Waltham, Massachusetts, 02454-9110, USA

    Yu Li

  3. Department of Biochemistry, Eötvös Loránd University, Pázmány P. s. 1/c, 1117, Budapest, Hungary

    Antal Blazsek, László Farkas & László Nyitray

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Supplementary information

41586_2003_BFnature01801_MOESM1_ESM.doc

Supplementary Information: The supplementary material provides the details of the expression, purification, crystallization, and structure determination (including a Table of X-ray data collection and refinement statistics) of the (leucine-zipper stabilized) 51-residue N-terminal fragment of scallop myosin subfragment-2. (DOC 30 kb)

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Li, Y., Brown, J., Reshetnikova, L. et al. Visualization of an unstable coiled coil from the scallop myosin rod. Nature 424, 341–345 (2003). https://doi.org/10.1038/nature01801

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