Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein
| dc.contributor.author | Crum, Mary Abou-Nader | |
| dc.contributor.author | Park, Jason M | |
| dc.contributor.author | Mulelu, Andani E | |
| dc.contributor.author | Sewell, Trevor B | |
| dc.contributor.author | Benedik, Michael J | |
| dc.date.accessioned | 2016-09-02T11:04:40Z | |
| dc.date.available | 2016-09-02T11:04:40Z | |
| dc.date.issued | 2015 | |
| dc.date.updated | 2016-09-02T11:03:41Z | |
| dc.description.abstract | The cyanide dihydratases from Bacillus pumilus and Pseudomonas stutzeri share high amino acid sequence similarity throughout except for their highly divergent C-termini. However, deletion or exchange of the C-termini had different effects upon each enzyme. Here we extended previous studies and investigated how the C-terminus affects the activity and stability of three nitrilases, the cyanide dihydratases from B. pumilus (CynDpum) and P. stutzeri (CynDstut) and the cyanide hydratase from Neurospora crassa. Enzymes in which the C-terminal residues were deleted decreased in both activity and thermostability with increasing deletion lengths. However, CynDstut was more sensitive to such truncation than the other two enzymes. A domain of the P. stutzeri CynDstut C-terminus not found in the other enzymes, 306GERDST311, was shown to be necessary for functionality and explains the inactivity of the previously described CynDstut-pum hybrid. This suggests that the B. pumilus C-terminus, which lacks this motif, may have specific interactions elsewhere in the protein, preventing it from acting in trans on a heterologous CynD protein. We identify the dimerization interface A-surface region 195–206 (A2) from CynDpum as this interaction site. However, this A2 region did not rescue activity in C-terminally truncated CynDstutΔ302 or enhance the activity of full-length CynDstut and therefore does not act as a general stability motif. | en_ZA |
| dc.identifier | http://dx.doi.org/10.1007/s00253-014-6335-x | |
| dc.identifier.apacitation | Crum, M. A., Park, J. M., Mulelu, A. E., Sewell, T. B., & Benedik, M. J. (2015). Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein. <i>Applied Microbiology and Biotechnology</i>, http://hdl.handle.net/11427/21658 | en_ZA |
| dc.identifier.chicagocitation | Crum, Mary Abou-Nader, Jason M Park, Andani E Mulelu, Trevor B Sewell, and Michael J Benedik "Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein." <i>Applied Microbiology and Biotechnology</i> (2015) http://hdl.handle.net/11427/21658 | en_ZA |
| dc.identifier.citation | Crum, M. A. N., Park, J. M., Mulelu, A. E., Sewell, B. T., & Benedik, M. J. (2015). Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein. Applied microbiology and biotechnology, 99(7), 3093-3102. | en_ZA |
| dc.identifier.issn | 0175-7598 | en_ZA |
| dc.identifier.ris | TY - Journal Article AU - Crum, Mary Abou-Nader AU - Park, Jason M AU - Mulelu, Andani E AU - Sewell, Trevor B AU - Benedik, Michael J AB - The cyanide dihydratases from Bacillus pumilus and Pseudomonas stutzeri share high amino acid sequence similarity throughout except for their highly divergent C-termini. However, deletion or exchange of the C-termini had different effects upon each enzyme. Here we extended previous studies and investigated how the C-terminus affects the activity and stability of three nitrilases, the cyanide dihydratases from B. pumilus (CynDpum) and P. stutzeri (CynDstut) and the cyanide hydratase from Neurospora crassa. Enzymes in which the C-terminal residues were deleted decreased in both activity and thermostability with increasing deletion lengths. However, CynDstut was more sensitive to such truncation than the other two enzymes. A domain of the P. stutzeri CynDstut C-terminus not found in the other enzymes, 306GERDST311, was shown to be necessary for functionality and explains the inactivity of the previously described CynDstut-pum hybrid. This suggests that the B. pumilus C-terminus, which lacks this motif, may have specific interactions elsewhere in the protein, preventing it from acting in trans on a heterologous CynD protein. We identify the dimerization interface A-surface region 195–206 (A2) from CynDpum as this interaction site. However, this A2 region did not rescue activity in C-terminally truncated CynDstutΔ302 or enhance the activity of full-length CynDstut and therefore does not act as a general stability motif. DA - 2015 DB - OpenUCT DP - University of Cape Town J1 - Applied Microbiology and Biotechnology LK - https://open.uct.ac.za PB - University of Cape Town PY - 2015 SM - 0175-7598 T1 - Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein TI - Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein UR - http://hdl.handle.net/11427/21658 ER - | en_ZA |
| dc.identifier.uri | http://hdl.handle.net/11427/21658 | |
| dc.identifier.uri | http://link.springer.com/article/10.1007/s00253-014-6335-x | |
| dc.identifier.vancouvercitation | Crum MA, Park JM, Mulelu AE, Sewell TB, Benedik MJ. Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein. Applied Microbiology and Biotechnology. 2015; http://hdl.handle.net/11427/21658. | en_ZA |
| dc.language | eng | en_ZA |
| dc.publisher | Springer Verlag | en_ZA |
| dc.publisher.institution | University of Cape Town | |
| dc.source | Applied Microbiology and Biotechnology | en_ZA |
| dc.source.uri | http://link.springer.com/journal/253 | |
| dc.subject.other | Cyanide | |
| dc.subject.other | dihydratase | |
| dc.subject.other | Nitrilase | |
| dc.subject.other | Cyanide | |
| dc.subject.other | Bioremediation | |
| dc.title | Probing C-terminal interactions of the Pseudomonas stutzeri cyanide-degrading CynD protein | en_ZA |
| dc.type | Journal Article | en_ZA |
| uct.type.filetype | Text | |
| uct.type.filetype | Image | |
| uct.type.publication | Research | en_ZA |
| uct.type.resource | Article | en_ZA |