Progranulin pp 51-64 | Cite as

Large-Scale Generation of Recombinant Granulin Peptides in E. coli

  • Dmitri TolkatchevEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1806)


Generating milligram quantities of correctly folded granulin molecules with properly formed disulfide bonds and biologically relevant activities may represent a considerable challenge. Here I describe a protocol for obtaining well-folded human granulins A, C, and F by expressing them as thioredoxin fusion proteins in Origami (DE3) Escherichia coli cells promoting disulfide bond formation in the cytoplasm environment. The thioredoxin tag is removed by proteolytic cleavage with enterokinase and granulins which are purified by reversed-phase HPLC. Well-folded disulfide species display lower retention time than misfolded species and therefore can be readily purified.

Key words

Granulin Progranulin Large-scale bacterial expression Disulfide bond Purification HPLC 


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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.The Gene and Linda Voiland School of Chemical Engineering and BioengineeringWashington State UniversityPullmanUSA

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