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Mendeley readers
Chapter title |
Large-Scale Generation of Recombinant Granulin Peptides in E. coli
|
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Chapter number | 4 |
Book title |
Progranulin
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Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8559-3_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8557-9, 978-1-4939-8559-3
|
Authors |
Dmitri Tolkatchev, Tolkatchev, Dmitri |
Abstract |
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. |
Mendeley readers
The data shown below were compiled from readership statistics for 2 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 2 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Bachelor | 1 | 50% |
Researcher | 1 | 50% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 1 | 50% |
Neuroscience | 1 | 50% |