Effect of signal peptide on stability and folding of Escherichia coli thioredoxin.
The signal peptide plays a key role in targeting and membrane insertion of secretory and membrane proteins in both prokaryotes and eukaryotes. In E. coli, recombinant proteins can be targeted to the periplasmic space by fusing naturally occurring signal sequences to their N-terminus. The model prote...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3646739?pdf=render |
| _version_ | 1818013358501134336 |
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| author | Pranveer Singh Likhesh Sharma S Rajendra Kulothungan Bharat V Adkar Ravindra Singh Prajapati P Shaik Syed Ali Beena Krishnan Raghavan Varadarajan |
| author_facet | Pranveer Singh Likhesh Sharma S Rajendra Kulothungan Bharat V Adkar Ravindra Singh Prajapati P Shaik Syed Ali Beena Krishnan Raghavan Varadarajan |
| author_sort | Pranveer Singh |
| collection | DOAJ |
| description | The signal peptide plays a key role in targeting and membrane insertion of secretory and membrane proteins in both prokaryotes and eukaryotes. In E. coli, recombinant proteins can be targeted to the periplasmic space by fusing naturally occurring signal sequences to their N-terminus. The model protein thioredoxin was fused at its N-terminus with malE and pelB signal sequences. While WT and the pelB fusion are soluble when expressed, the malE fusion was targeted to inclusion bodies and was refolded in vitro to yield a monomeric product with identical secondary structure to WT thioredoxin. The purified recombinant proteins were studied with respect to their thermodynamic stability, aggregation propensity and activity, and compared with wild type thioredoxin, without a signal sequence. The presence of signal sequences leads to thermodynamic destabilization, reduces the activity and increases the aggregation propensity, with malE having much larger effects than pelB. These studies show that besides acting as address labels, signal sequences can modulate protein stability and aggregation in a sequence dependent manner. |
| first_indexed | 2024-04-14T06:32:11Z |
| format | Article |
| id | doaj.art-f3bb9abef7044244b43609a4d4a9b2a2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-14T06:32:11Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f3bb9abef7044244b43609a4d4a9b2a22022-12-22T02:07:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0185e6344210.1371/journal.pone.0063442Effect of signal peptide on stability and folding of Escherichia coli thioredoxin.Pranveer SinghLikhesh SharmaS Rajendra KulothunganBharat V AdkarRavindra Singh PrajapatiP Shaik Syed AliBeena KrishnanRaghavan VaradarajanThe signal peptide plays a key role in targeting and membrane insertion of secretory and membrane proteins in both prokaryotes and eukaryotes. In E. coli, recombinant proteins can be targeted to the periplasmic space by fusing naturally occurring signal sequences to their N-terminus. The model protein thioredoxin was fused at its N-terminus with malE and pelB signal sequences. While WT and the pelB fusion are soluble when expressed, the malE fusion was targeted to inclusion bodies and was refolded in vitro to yield a monomeric product with identical secondary structure to WT thioredoxin. The purified recombinant proteins were studied with respect to their thermodynamic stability, aggregation propensity and activity, and compared with wild type thioredoxin, without a signal sequence. The presence of signal sequences leads to thermodynamic destabilization, reduces the activity and increases the aggregation propensity, with malE having much larger effects than pelB. These studies show that besides acting as address labels, signal sequences can modulate protein stability and aggregation in a sequence dependent manner.http://europepmc.org/articles/PMC3646739?pdf=render |
| spellingShingle | Pranveer Singh Likhesh Sharma S Rajendra Kulothungan Bharat V Adkar Ravindra Singh Prajapati P Shaik Syed Ali Beena Krishnan Raghavan Varadarajan Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. PLoS ONE |
| title | Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. |
| title_full | Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. |
| title_fullStr | Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. |
| title_full_unstemmed | Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. |
| title_short | Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. |
| title_sort | effect of signal peptide on stability and folding of escherichia coli thioredoxin |
| url | http://europepmc.org/articles/PMC3646739?pdf=render |
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