Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy
Staphylococcus aureus hibernation-promoting factor (SaHPF) is a 22.2 kDa translation factor which under stress conditions (under amino acid starvation or antibiotic pressure) binds with the ribosome and inactivates it, thereby ensuring cell survival under stress. There are many problems with crystal...
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| Format: | Article |
| Language: | English |
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Kazan Federal University
2017-06-01
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| Series: | Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki |
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| Online Access: | http://kpfu.ru/portal/docs/F1383402460/159_2_est_12.pdf |
| _version_ | 1819034701690568704 |
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| author | R.Kh. Ayupov K.S. Usachev I.Sh. Khusainov B. Kieffer M.M. Yusupov |
| author_facet | R.Kh. Ayupov K.S. Usachev I.Sh. Khusainov B. Kieffer M.M. Yusupov |
| author_sort | R.Kh. Ayupov |
| collection | DOAJ |
| description | Staphylococcus aureus hibernation-promoting factor (SaHPF) is a 22.2 kDa translation factor which under stress conditions (under amino acid starvation or antibiotic pressure) binds with the ribosome and inactivates it, thereby ensuring cell survival under stress. There are many problems with crystallization of this protein which still remain unsolved. Therefore, its analysis by NMR spectroscopy is of great interest. In this paper, we have described expression, purification, and NMR analysis of 13C/15N-labeled SaHPF protein and showed that it is present in a dimeric form in the solution. Notably, two types of signals in the NMR spectra have been observed: with weak intensity and high dispersion from N-terminal domain; with high intensity but low dispersion from a flexible loop between domains. No signals from C-terminal domain have been observed in the NMR spectra, which may indicate possible dimerization of this part of the protein. Protein dimerization has been also detected by the method of electrophoresis under native conditions. |
| first_indexed | 2024-12-21T07:37:55Z |
| format | Article |
| id | doaj.art-f4cc4fa511b34f8daba07a535c8c44c1 |
| institution | Directory Open Access Journal |
| issn | 2542-064X 2500-218X |
| language | English |
| last_indexed | 2024-12-21T07:37:55Z |
| publishDate | 2017-06-01 |
| publisher | Kazan Federal University |
| record_format | Article |
| series | Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki |
| spelling | doaj.art-f4cc4fa511b34f8daba07a535c8c44c12022-12-21T19:11:24ZengKazan Federal UniversityUčënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki2542-064X2500-218X2017-06-011592332341Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR SpectroscopyR.Kh. Ayupov0K.S. Usachev1I.Sh. Khusainov2B. Kieffer3M.M. Yusupov4Kazan Federal University, Kazan, 420008 RussiaKazan Federal University, Kazan, 420008 RussiaKazan Federal University, Kazan, 420008 Russia; Institute of Genetics and Molecular and Cellular Biology (IGBMC), Illkirch-Graffenstaden, 67400 FranceInstitute of Genetics and Molecular and Cellular Biology (IGBMC), Illkirch-Graffenstaden, 67400 FranceKazan Federal University, Kazan, 420008 RussiaStaphylococcus aureus hibernation-promoting factor (SaHPF) is a 22.2 kDa translation factor which under stress conditions (under amino acid starvation or antibiotic pressure) binds with the ribosome and inactivates it, thereby ensuring cell survival under stress. There are many problems with crystallization of this protein which still remain unsolved. Therefore, its analysis by NMR spectroscopy is of great interest. In this paper, we have described expression, purification, and NMR analysis of 13C/15N-labeled SaHPF protein and showed that it is present in a dimeric form in the solution. Notably, two types of signals in the NMR spectra have been observed: with weak intensity and high dispersion from N-terminal domain; with high intensity but low dispersion from a flexible loop between domains. No signals from C-terminal domain have been observed in the NMR spectra, which may indicate possible dimerization of this part of the protein. Protein dimerization has been also detected by the method of electrophoresis under native conditions.http://kpfu.ru/portal/docs/F1383402460/159_2_est_12.pdfSaHPFStaphylococcus aureusNMRribosomeantibiotictranslation factor |
| spellingShingle | R.Kh. Ayupov K.S. Usachev I.Sh. Khusainov B. Kieffer M.M. Yusupov Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki SaHPF Staphylococcus aureus NMR ribosome antibiotic translation factor |
| title | Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy |
| title_full | Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy |
| title_fullStr | Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy |
| title_full_unstemmed | Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy |
| title_short | Expression and Purification of HPF Protein from Staphylococcus aureus and Analysis of Its Structure by the Method of NMR Spectroscopy |
| title_sort | expression and purification of hpf protein from staphylococcus aureus and analysis of its structure by the method of nmr spectroscopy |
| topic | SaHPF Staphylococcus aureus NMR ribosome antibiotic translation factor |
| url | http://kpfu.ru/portal/docs/F1383402460/159_2_est_12.pdf |
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