Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity.
Inteins catalyze a protein splicing reaction to excise the intein from a precursor protein and join the flanking sequences (exteins) with a peptide bond. In a split intein, the intein fragments (I(N) and I(C)) can reassemble non-covalently to catalyze a trans-splicing reaction that joins the exteins...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3443213?pdf=render |
| _version_ | 1818319407135326208 |
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| author | Huiling Song Qing Meng Xiang-Qin Liu |
| author_facet | Huiling Song Qing Meng Xiang-Qin Liu |
| author_sort | Huiling Song |
| collection | DOAJ |
| description | Inteins catalyze a protein splicing reaction to excise the intein from a precursor protein and join the flanking sequences (exteins) with a peptide bond. In a split intein, the intein fragments (I(N) and I(C)) can reassemble non-covalently to catalyze a trans-splicing reaction that joins the exteins from separate polypeptides. An atypical split intein having a very small I(N) and a large I(C) is particularly useful for joining synthetic peptides with recombinant proteins, which can be a generally useful method of introducing site-specific chemical labeling or modifications into proteins. However, a large I(C) derived from an Ssp DnaX intein was found recently to undergo spontaneous C-cleavage, which raised questions regarding its structure-function and ability to trans-splice. Here, we show that this I(C) could undergo trans-splicing in the presence of I(N), and the trans-splicing activity completely suppressed the C-cleavage activity. We also found that this I(C) could trans-splice with small I(N) sequences derived from two other inteins, showing a cross-reactivity of this atypical split intein. Furthermore, we found that this I(C) could trans-splice even when the I(N) sequence was embedded in a nearly complete intein sequence, suggesting that the small I(N) could project out of the central pocket of the intein to become accessible to the I(C). Overall, these findings uncovered a new atypical split intein that can be valuable for peptide-protein trans-splicing, and they also revealed an interesting structural flexibility and cross-reactivity at the active site of this intein. |
| first_indexed | 2024-12-13T10:08:37Z |
| format | Article |
| id | doaj.art-d2821ac2c97740e6808a30eb6e882315 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T10:08:37Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-d2821ac2c97740e6808a30eb6e8823152022-12-21T23:51:30ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0179e4535510.1371/journal.pone.0045355Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity.Huiling SongQing MengXiang-Qin LiuInteins catalyze a protein splicing reaction to excise the intein from a precursor protein and join the flanking sequences (exteins) with a peptide bond. In a split intein, the intein fragments (I(N) and I(C)) can reassemble non-covalently to catalyze a trans-splicing reaction that joins the exteins from separate polypeptides. An atypical split intein having a very small I(N) and a large I(C) is particularly useful for joining synthetic peptides with recombinant proteins, which can be a generally useful method of introducing site-specific chemical labeling or modifications into proteins. However, a large I(C) derived from an Ssp DnaX intein was found recently to undergo spontaneous C-cleavage, which raised questions regarding its structure-function and ability to trans-splice. Here, we show that this I(C) could undergo trans-splicing in the presence of I(N), and the trans-splicing activity completely suppressed the C-cleavage activity. We also found that this I(C) could trans-splice with small I(N) sequences derived from two other inteins, showing a cross-reactivity of this atypical split intein. Furthermore, we found that this I(C) could trans-splice even when the I(N) sequence was embedded in a nearly complete intein sequence, suggesting that the small I(N) could project out of the central pocket of the intein to become accessible to the I(C). Overall, these findings uncovered a new atypical split intein that can be valuable for peptide-protein trans-splicing, and they also revealed an interesting structural flexibility and cross-reactivity at the active site of this intein.http://europepmc.org/articles/PMC3443213?pdf=render |
| spellingShingle | Huiling Song Qing Meng Xiang-Qin Liu Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity. PLoS ONE |
| title | Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity. |
| title_full | Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity. |
| title_fullStr | Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity. |
| title_full_unstemmed | Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity. |
| title_short | Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity. |
| title_sort | protein trans splicing of an atypical split intein showing structural flexibility and cross reactivity |
| url | http://europepmc.org/articles/PMC3443213?pdf=render |
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