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Crystallographic Reconstruction of the Acrosomal Process from Limulus Polyphemus Sperm

Published online by Cambridge University Press:  02 July 2020

M. B. Sherman ,
J. Jakana ,
S. Sun ,
P. Matsudaira ,
W. Chiu  and
M. F. Schmid
M. B. Sherman
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Dept. of Biochemistry, Baylor College of Medicine, Houston, TX, 77030
J. Jakana
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Dept. of Biochemistry, Baylor College of Medicine, Houston, TX, 77030
S. Sun
Affiliation:
Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02142
P. Matsudaira
Affiliation:
Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02142
W. Chiu
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Dept. of Biochemistry, Baylor College of Medicine, Houston, TX, 77030
M. F. Schmid
Affiliation:
National Center for Macromolecular Imaging and Verna and Marrs McLean Dept. of Biochemistry, Baylor College of Medicine, Houston, TX, 77030
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Extract

The acrosomal process is an intracellular quasi-crystalline organelle in the head of the sperm of the horseshoe crab Limulus polyphemus. It consists of 100 - 130 actin-scruin filaments packed together in a pseudo-hexagonal lattice2 and is up to 60 (im long with a diameter of 0.1 μm. Scruin-scruin interactions are responsible for cross-linking the actin filaments together in the bundle. The goal of the current study is to reveal interfilament interactions in the bundle. We have taken tilt series images in the electron microscope3 to reconstruct its three-dimensional structure without assuming helical symmetry.

The acrosomal process was purified as described.34 Bundles were embedded in vitreous ice over holes on a holey carbon film on copper grids. The specimen was kept at -167°C in a JEOL 4000EX electron microscope operating at 400 kV. Straight 6-10 (im long bundles were found using a TV-rate CCD camera in defocused diffraction mode.

Type
High Resolution Protein Structures from Electron Crystallography
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 460 - 461
Copyright
Copyright © Microscopy Society of America

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References

References:

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7. Grant support NIH-RR02250, NSF-BIR9412521, NSF-BIR9500098 and NIH-GM52703.Google Scholar