Prediction of hot spots in dimer interface of green fluorescent protein

Abstract

Wild-type green fluorescent protein (GFP) has a weak dimerization tendency at higher concentrations. Currently there are still a limited number of known mutated residues for the dimer interface breaking. The crystal structure of the dimer provides vital background on neighbouring residue interactions. Here well-established hot spot prediction methods, such as HotPoint, KFC2, PredHS and Robetta, were adopted to predict residues critical for dimer stabilization, i.e. hot spots. Among 17 residues appeared on the dimer interface of GFP, 10 residues were predicted as hot spots by at least one method. Some of these computational hot spots were verified with existing site-directed mutagenesis results, and the remains of the predicted residues can be candidates for further validation. Furthermore, the structural feature such as relative accessibility was found significantly different between computational hot spots and non-hot spots.