| Title |
Expression and characterization of human proinsulin fused to thioredoxin in Escherichia coli
|
|---|---|
| Published in |
Applied Microbiology and Biotechnology, December 2011
|
| DOI | 10.1007/s00253-011-3721-5 |
| Pubmed ID | |
| Authors |
Aldana Trabucchi, Luciano Lucas Guerra, Natalia Inés Faccinetti, Ruben Francisco Iacono, Edgardo Poskus, Silvina Noemí Valdez |
| Abstract |
Native proinsulin (PI) belongs to the class of the difficult-to-express proteins in Escherichia coli. Problems mainly arise due to its high proteolytic decay and troubles to reproduce the native disulphide pattern. In the present study, human PI was produced in E. coli as a fusion thioredoxin protein (Trx-PI). Such chimeric protein was obtained from the intracellular soluble fraction, and it was purified in one step by affinity chromatography on immobilized phenylarsine oxide. Trx-PI was also recovered from inclusion bodies and purified by anion exchange chromatography. The product identity and integrity were verified by mass analysis (22,173.5 Da) and mapping with Staphylococcus aureus V8 protease. Native PI folding was evaluated by biochemical and also by immunochemical analysis using specific sera from PI antibody-positive diabetic patients that recognise conformational discontinue epitopes. Dose-response curves showed identity between standard PI and Trx-PI. Moreover, surface plasmon resonance technique verified the correct conformation of the recombinant protein. The biochemical and immunochemical assays demonstrated the integrity of the chimera and the epitopes involved in the interaction with antibodies. In conclusion, it was possible to obtain with high-yield purified human PI as a fusion protein in E. coli and useful for analytical purposes. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 25 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Switzerland | 1 | 4% |
| Unknown | 24 | 96% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 5 | 20% |
| Student > Bachelor | 3 | 12% |
| Researcher | 3 | 12% |
| Student > Doctoral Student | 2 | 8% |
| Professor | 1 | 4% |
| Other | 3 | 12% |
| Unknown | 8 | 32% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 7 | 28% |
| Medicine and Dentistry | 2 | 8% |
| Nursing and Health Professions | 1 | 4% |
| Biochemistry, Genetics and Molecular Biology | 1 | 4% |
| Veterinary Science and Veterinary Medicine | 1 | 4% |
| Other | 4 | 16% |
| Unknown | 9 | 36% |
Attention Score in Context
This research output has an Altmetric Attention Score of 4. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 06 June 2013.
All research outputs
#7,480,940
of 24,119,703 outputs
Outputs from Applied Microbiology and Biotechnology
#2,515
of 8,034 outputs
Outputs of similar age
#65,755
of 247,309 outputs
Outputs of similar age from Applied Microbiology and Biotechnology
#37
of 82 outputs
Altmetric has tracked 24,119,703 research outputs across all sources so far. This one has received more attention than most of these and is in the 68th percentile.
So far Altmetric has tracked 8,034 research outputs from this source. They receive a mean Attention Score of 4.3. This one has gotten more attention than average, scoring higher than 67% of its peers.
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We're also able to compare this research output to 82 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 53% of its contemporaries.