Split-superpositive GFP is among the most accessed articles

split-GFP reassembly is more efficient than previously reported systems

Posted: July 26th, 2012

A recent paper from the McNaughton lab, titled “Split-superpositive GFP reassembly is a fast, efficient, and robust method for detecting protein–protein interactions in vivo” has been cited as one of the most accessed journal articles in Molecular BioSystems during the month of June (http://blogs.rsc.org/mb/category/top-ten/). Molecular BioSystems publishes high-quality interdisciplinary research with a particular focus on the interface between chemical biology and the -omic sciences and systems biology. Split-Green Fluorescent Protein (GFP) reassembly is an operationally simple in vivo technique used to identify and study interactions involving proteins and/or peptides.  However, the instability of split-GFP fragments, and their susceptibility to aggregation, place limitations on the broader use of this method. Researchers Brett Blakeley and Alex Chapman hypothesized that re-engineered split-GFP fragments with high net charge may increase their solubility and make them resistant to aggregation, thus overcoming barriers to interaction-dependent split-GFP reassemly. Blakeley and Chapman showed that a split-superpositive GFP (split-spGFP) variant reassembles faster and more efficiently than previously reported systems.  In addition, interaction-dependent split-spGFP reassembly is efficient at physiological temperature.  The improved properties of split-spGFP reassembly make this reporter system ideal for identifying and studying interactions involving proteins and/or peptides, and may be particularly useful for identifying or studying interactions involving proteins or peptides that are themselves susceptible to aggregation.