Cryoelectron tomography: Implications for actin cytoskeleton research

Weber, Igor (2005) Cryoelectron tomography: Implications for actin cytoskeleton research. Croatica Chemica Acta, 78 (3). pp. 325-331. ISSN 0011-1643

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Abstract

Disclosing undistorted spatial organization of the actin cytoskeleton with molecular resolution is fundamental for understanding the cellular ultrastructure. Whereas important insights into the architecture of microfilament networks have been gained by negative staining, critical point drying, and freeze-fracturing methods, it is cryoelectron tomography that provides, for the first time, a comprehensive three-dimensional view of the intact actin cytoskeleton in situ. In particular, topological relationships such as microfilament three-dimensional proximity, angles at filament branching points, and modes of microfilament interaction with the plasma membrane can be visualized with an unprecedented accuracy using this technique. Further improvements are expected to bring the resolution into the realm of 2-3 nm, where automatic pattern-recognition methods can be applied to identify actin-binding complexes. Combining cryoelectron tomography with ultra-fine immunolabeling and high-resolution fluorescence microscopy will make it possible to correlate structural data on the nanometer scale with molecular specificity and dynamical information.

Item Type:	Article
Uncontrolled Keywords:	actin; cytoskeleton; cryofixation; electron microscopy; tomography; electron-microscopy; macromolecular complexes; 3-dimensional structure; dynamic organization; arp2/3 complex; cells; dictyostelium; system; model
Subjects:	NATURAL SCIENCES > Chemistry
Divisions:	Division of Molecular Biology
Projects:	Project title Project leader Project code Project type Struktura i funkcija plastida i citoskeleta Nikola Ljubešić 0098073 MZOS
Depositing User:	Igor Weber
Date Deposited:	31 Oct 2013 13:12
URI:	http://fulir.irb.hr/id/eprint/881

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