Tagging recombinant proteins to enhance solubility and aid purification.
Protein fusion technology has enormously facilitated the efficient production and purification of individual recombinant proteins. The use of genetically engineered affinity and solubility-enhancing polypeptide "tags" has increased greatly in recent years and there now exists a considerable repertoire of these that can be used to solve issues related to the expression, stability, solubility, folding, and purification of their fusion partner. In the case of large-scale proteomic studies, the development of purification procedures tailored to individual proteins is not practicable, and affinity tags have therefore become indispensable tools for structural and functional proteomic initiatives that involve the expression of many proteins in parallel. Here, the rationale and applications of a range of established and more recently developed solubility-enhancing and affinity tags are outlined.
Walls
Dermot
D
School of Biotechnology and National Centre for Sensor Research, Dublin City University, Dublin, Ireland. dermot.walls@dcu.ie
Loughran
Sinéad T
ST
D000818:Animals; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D058619:Maltose-Binding Proteins / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D025842:SUMO-1 Protein / Q000235:genetics; D012995:Solubility
D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; D005982:Glutathione Transferase