Positional effects of fusion partners on the yield and solubility of MBP fusion proteins

Protein Expression and Purification
Sreejith Raran-KurussiD S Waugh

Abstract

Escherichia coli maltose-binding protein (MBP) is exceptionally effective at promoting the solubility of its fusion partners. However, there are conflicting reports in the literature claiming that (1) MBP is an effective solubility enhancer only when it is joined to the N-terminus of an aggregation-prone passenger protein, and (2) MBP is equally effective when fused to either end of the passenger. Here, we endeavor to resolve this controversy by comparing the solubility of a diverse set of MBP fusion proteins that, unlike those analyzed in previous studies, are identical in every way except for the order of the two domains. The results indicate that fusion proteins with an N-terminal MBP provide an excellent solubility advantage along with more robust expression when compared to analogous fusions in which MBP is the C-terminal fusion partner. We find that only intrinsically soluble passenger proteins (i.e., those not requiring a solubility enhancer) are produced as soluble fusions when they precede MBP. We also report that even subtle differences in inter-domain linker sequences can influence the solubility of fusion proteins.

References

Aug 19, 1999·Protein Science : a Publication of the Protein Society·R B Kapust, D S Waugh
Sep 26, 2001·Protein Expression and Purification·Y NominéG Travé
Jun 21, 2002·Biochemical and Biophysical Research Communications·Rachel B KapustDavid S Waugh
Jun 18, 2005·Biochemical and Biophysical Research Communications·Pierre DouetteFrancis E Sluse
Sep 20, 2005·Protein Expression and Purification·Sreedevi Nallamsetty, David S Waugh
Jun 20, 2006·Current Opinion in Biotechnology·Dominic Esposito, Deb K Chatterjee
Jun 27, 2006·Protein Expression and Purification·Mareike KurzPawel Listwan
Sep 23, 2009·Acta Crystallographica. Section D, Biological Crystallography·George T LountosDavid S Waugh
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Stephen N HewittWesley C Van Voorhis

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Citations

Dec 20, 2015·Protein Science : a Publication of the Protein Society·David S Waugh
Jul 7, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Bostjan KobeSimon J Williams
Apr 24, 2016·Analytical Biochemistry·Sreejith Raran-Kurussi, David S Waugh
Apr 27, 2012·PloS One·Marc GuerineauJan Palecek
Jul 5, 2016·Frontiers in Molecular Biosciences·Zhenyu HaoAdrian Goldman
Apr 2, 2005·Protein Expression and Purification·Ashima MitraSiddhartha P Sarma
Nov 5, 2016·Archives of Biochemistry and Biophysics·Dinesh K YadavNarendra Tuteja
Jan 30, 2018·Biotechnology and Applied Biochemistry·Svetlana P IkonomovaAmy J Karlsson
Nov 11, 2018·Bioscience Reports·Lalith K ChagantiKakoli Bose
Jan 4, 2018·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Seung Won YangBaik Lin Seong
Oct 20, 2018·Biotechnology for Biofuels·Lijun YeXueli Zhang
Oct 23, 2019·Scientific Reports·Maria S YurkovaAlexey N Fedorov
Jul 24, 2021·International Journal of Biological Macromolecules·Michal NemergutErik Sedlák

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