Stroupe, M.E., Xu, C., Goode, B.L., Grigorieff, N. (2009). Actin filament labels for localizing protein components in large complexes viewed by electron microscopy.  RNA 15(2): 244--248.

FBrf0215711

Research paper

Localizing specific components in three-dimensional reconstructions of protein complexes visualized in an electron microscope increases the scientific value of those structures. Subunits are often identified within the complex by labeling; however, unless the label produces directly visible features, it must be detected by computational comparison with unlabeled complex. To bypass this step, we generated a cloneable tag from the actin-nucleating protein Spire that produces a directly visible "pointer" to the subunit after actin polymerization. We have used this new label to identify the intron of the C complex spliceosome to its small domain by fusing the 10 kDa Spire moiety to the affinity label that binds recombinant stem loops in the pre-mRNA substrate and assembling an actin filament on the particle.

PMC2648703 (PMC) (EuropePMC)