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Electron Crystallographic Structure of the Limulus Acrosomal Bundle at 20 Å Resolution

Published online by Cambridge University Press:  02 July 2020

Michael B. Sherman ,
Guichy Waller ,
Paul Matsudaira ,
Wah Chiu  and
Michael F. Schmid
Michael B. Sherman
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX77030
Guichy Waller
Affiliation:
Whitehead Institute for Biomedical Research and Department of Biology and Division of Bioengineering and Environmental Health, Massachusetts Institute of Technology, Cambridge, MA02142
Paul Matsudaira
Affiliation:
Whitehead Institute for Biomedical Research and Department of Biology and Division of Bioengineering and Environmental Health, Massachusetts Institute of Technology, Cambridge, MA02142 Department of Biology and Division of Bioengineering and Environmental Health, Massachusetts Institute of Technology, Cambridge, MA02142
Wah Chiu
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX77030
Michael F. Schmid
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX77030
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Extract

Limulus sperm contains a dynamic macromolecular structure that rapidly extends a 50-μum process called the true discharge. The core of this structure is a bundle of ordered filaments composed of a complex of actin, scruin and calmodulin. We have shown that small segments along the bundle can be treated as single crystals with a unit cell spacing of 144 × 144 × 766 Å. A tomographic reconstruction of the bundle was done from multiple tilt series of images to ∼40 Å resolution. To extend the structural determination of the bundle at a higher resolution, we have used electron crystallographic analysis of single images of the bundles preserved in vitreous ice. Furthermore, we did not employ any helical or crystallographic symmetry (other than PI) in the reconstruction procedure.

Acrosomal bundles from Limulus sperm cells were purified as described earlier. Images of frozen hydrated bundles were taken at 40,000x EM magnification in a JEOL 4000EX electron cryomicroscope using an electron dose of 6 - 10 electrons/Å2.

Type
Electron Cryomicroscopy of Macromolecules
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 242 - 243
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Schmid, M.F. et al. (1991) J Mol Biol 221, 711–725.CrossRefGoogle Scholar
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5. This research was supported by NIH grant RR02250.Google Scholar