title: Unrelated solubility-enhancing fusion partners MBP and NusA utilize a similar mode of action.
abstract: The tendency of recombinant proteins to accumulate in the form of insoluble aggregates in Escherichia coli is a major hindrance to their overproduction. One of the more effective approaches to circumvent this problem is to use translation fusion partners {solubility-enhancers (SEs)}. E. coli maltose-binding protein (MBP) and N-utilization substance A (NusA) are arguably the most effective solubilizing agents that have been discovered so far. Here, we show that although these two proteins are structurally, functionally, and physicochemically distinct, they influence the solubility and folding of their fusion partners in a very similar manner. These SEs act as "holdases" that prevent the aggregation of their fusion partners. Subsequent folding of the passenger proteins, when it occurs, is either spontaneous or chaperone-mediated.
authors.affiliation: Protein Engineering Section, Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, Maryland.
authors.forename: Sreejith
authors.initials: S
authors.lastname: Raran-Kurussi
authors.forename: David S
authors.initials: DS
authors.lastname: Waugh
mesh_terms: D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D002479:Inclusion Bodies; D058619:Maltose-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D010445:Peptide Elongation Factors / Q000737:chemistry* / Q000378:metabolism*; D015202:Protein Engineering; D058849:Protein Refolding; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism*; D012995:Solubility; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D035602:Transcriptional Elongation Factors
chemical_list: D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D010445:Peptide Elongation Factors; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C068727:nusA protein, E coli
keywords: MBP; NusA; chaperone; fusion partners; inclusion bodies; solubility-enhancers