Precise mapping of subunits in multiprotein complexes by a versatile electron microscopy label.

Dirk Flemming, Karsten Thierbach, Philipp Stelter, Bettina Böttcher, Ed Hurt

Biochemie-Zentrum der Universität Heidelberg, Heidelberg, Germany.

Nature structural & molecular biology 2010 Jun

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Positional knowledge of subunits within multiprotein assemblies is crucial for understanding their function. The topological analysis of protein complexes by electron microscopy has undergone impressive development, but analysis of the exact positioning of single subunits has lagged behind. Here we have developed a clonable approximately 80-residue tag that, upon attachment to a target protein, can recruit a structurally prominent electron microscopy label in vitro. This tag is readily visible on single particles and becomes exceptionally distinct after image processing and classification. Thus, our method is applicable for the exact topological mapping of subunits in macromolecular complexes.

Citation

Dirk Flemming, Karsten Thierbach, Philipp Stelter, Bettina Böttcher, Ed Hurt. Precise mapping of subunits in multiprotein complexes by a versatile electron microscopy label. Nature structural & molecular biology. 2010 Jun;17(6):775-8