Hostname: page-component-cd9895bd7-7cvxr Total loading time: 0 Render date: 2024-12-28T03:55:57.504Z Has data issue: false hasContentIssue false
  • English
  • Français

New Strategy for Crystallographic Reconstruction of Biological Bundle-Like Structures

Published online by Cambridge University Press:  02 July 2020

M.B. Sherman ,
J. Jakana ,
S. Sunt ,
p. Matsudaira ,
W. Chiu  and
M.F. Schmid
M.B. Sherman
Affiliation:
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, One Baylor Plaza, Houston, TX77030;
J. Jakana
Affiliation:
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, One Baylor Plaza, Houston, TX77030;
S. Sunt
Affiliation:
The Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02142
p. Matsudaira
Affiliation:
The Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02142
W. Chiu
Affiliation:
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, One Baylor Plaza, Houston, TX77030;
M.F. Schmid
Affiliation:
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, One Baylor Plaza, Houston, TX77030;
Get access

Extract

The acrosomal bundle is an intracellular quasi-crystalline organelle in the head of the sperm of Limulus polyphemus . It is a long (up to 60 μm) straight bundle ∼1000 Å in diameter1 consisting of two major proteins in a 1:1 stoichiometric ratio: actin (42 kDa) and scruin (102 kDa) with a minor calmodulin-like protein (14 kDa) presumably bound to scruin molecule. Previous helical reconstructions of single filaments in the bundle showed actin-scruin interactions. We recorded tilt series from single bundles in a 400 kV cryomicroscope and have developed a novel crystallographic technique to reconstruct a unit cell from the bundle as a whole to reveal interfilament interactions.

Acrosomal bundles were purified as described elsewhere and embedded in vitreous ice over holes on a holey carbon film. Tilt series images were collected in a JEOL 4000EX electron cryo-microscope at 400 kV using 10,000x microscope magnification. Total dose was ∼16-18 electrons/Å per series covering a tilt range of ± 60° with 5° angular increment.

Type
Electron Crystallography; the Electron Phase Problem
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 1035 - 1036
Copyright
Copyright © Microscopy Society of America 1997

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1.Tilney., L.G.. J. Cell Biol. 64 ( 1975) 28910.1083/jcb.64.2.289CrossRefGoogle Scholar
2.Schmid, M.F.et al.. W. J. Cell Biol. 124(1994) 34110.1083/jcb.124.3.341CrossRefGoogle Scholar
3.Owen, C. and DeRosier, D.. J. Cell Biol. 123 (1993) 33710.1083/jcb.123.2.337CrossRefGoogle Scholar
4.Schmid, M.F.et al.J. Mol. Biol. 221 (1991) 71110.1016/0022-2836(91)80082-6CrossRefGoogle Scholar
5.Schmid, M.F.et al.J. Struct. Biol. 108 (1992) 62.10.1016/1047-8477(92)90007-WCrossRefGoogle Scholar
6. This research has been supported by NIH grant RR02250.Google Scholar