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Two-dimensional crystallization of brush border myosin I
H Celia1, J D Jontes, M Whittaker
1Department of Cell Biology MB25, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California, 92037, USA. milligan@scripps.edu
Journal of Structural Biology
|November 1, 1996
Summary
Brush border myosin-I (BBMI) forms two-dimensional crystals by binding to negatively charged lipids. This advance enables structural analysis of BBMI, crucial for intestinal epithelial cell function.
Area of Science:
- Biochemistry
- Cell Biology
- Structural Biology
Background:
- Brush border myosin-I (BBMI) is a single-headed unconventional myosin.
- BBMI is located in intestinal epithelial cell microvilli, linking actin filaments to the plasma membrane.
- BBMI's carboxy-terminal domain, rich in basic amino acids, mediates its association with anionic phospholipids.
Purpose of the Study:
- To exploit BBMI's affinity for negatively charged lipids.
- To form two-dimensional (2D) crystals of BBMI suitable for structural analysis.
- To determine the structural characteristics of BBMI 2D crystals.
Main Methods:
- Utilizing the natural affinity of BBMI for anionic phospholipids.
- Forming two-dimensional (2D) crystals of BBMI.
- Employing electron crystallographic techniques for structural analysis.
- Calculating projection maps from negatively stained crystal images.
Main Results:
- Successfully formed 2D crystals of BBMI.
- The crystals belong to space groups p22121 or p2.
- Projection maps were calculated to a resolution of 20 Å.
- The asymmetric unit was found to be identical in both crystal types.
Conclusions:
- BBMI's interaction with anionic lipids facilitates 2D crystal formation.
- These 2D crystals are suitable for high-resolution structural studies.
- The structural data provides insights into BBMI's function in intestinal epithelial cells.