Solubility-enhancing proteins MBP and NusA play a passive role in the folding of their fusion partners.
It is well established that certain highly soluble proteins have the ability to enhance the solubility of their fusion partners. However, very little is known about how different solubility enhancers compare in terms of their ability to promote the proper folding of their passenger proteins. We compared the ability of two well-known solubility enhancers, Escherichia coli maltose-binding protein (MBP) and N utilization substance A (NusA), to improve the solubility and promote the proper folding of a variety of passenger proteins that are difficult to solubilize. We used an intracellular processing system to monitor the solubility of these passenger proteins after they were cleaved from MBP and NusA by tobacco etch virus protease. In addition, the biological activity of some fusion proteins was compared to serve as a more quantitative indicator of native structure. The results indicate that MBP and NusA have comparable solubility-enhancing properties. Little or no difference was observed either in the solubility of passenger proteins after intracellular processing of the MBP and NusA fusion proteins or in the biological activity of solubilized passenger proteins, suggesting that the underlying mechanism of solubility enhancement is likely to be similar for both the proteins, and that they play a passive role rather than an active one in the folding of their fusion partners.
Nallamsetty
Sreedevi
S
Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, P.O. Box B, Frederick, MD, USA.
Waugh
David S
DS
D002352:Carrier Proteins / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry; D005822:Genetic Vectors / Q000737:chemistry; D058619:Maltose-Binding Proteins; D010445:Peptide Elongation Factors / Q000737:chemistry*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012995:Solubility; D013997:Time Factors; D014157:Transcription Factors / Q000737:chemistry*; D035602:Transcriptional Elongation Factors
D002352:Carrier Proteins; 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