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Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography
09:23

Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography

Published on: October 30, 2010

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
PubMed

Insights

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.

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