Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein.
Human respiratory syncytial virus (HRSV) has a nonsegmented negative-stranded RNA genome which is encapsidated by the HRSV nucleocapsid protein (HRSVN) that is essential for viral replication. HRSV is a common cause of respiratory infection in infants, yet no effective antiviral drugs to combat it a...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2008
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| _version_ | 1797069335497277440 |
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| author | El Omari, K Scott, K Dhaliwal, B Ren, J Abrescia, N Budworth, J Lockyer, M Powell, K Hawkins, A Stammers, D |
| author_facet | El Omari, K Scott, K Dhaliwal, B Ren, J Abrescia, N Budworth, J Lockyer, M Powell, K Hawkins, A Stammers, D |
| author_sort | El Omari, K |
| collection | OXFORD |
| description | Human respiratory syncytial virus (HRSV) has a nonsegmented negative-stranded RNA genome which is encapsidated by the HRSV nucleocapsid protein (HRSVN) that is essential for viral replication. HRSV is a common cause of respiratory infection in infants, yet no effective antiviral drugs to combat it are available. Recent data from an experimental anti-HRSV compound, RSV-604, indicate that HRSVN could be the target site for drug action. Here, the expression, purification and preliminary data collection of decameric HRSVN as well as monomeric N-terminally truncated HRSVN mutants are reported. Two different crystal forms of full-length selenomethionine-labelled HRSVN were obtained that diffracted to 3.6 and approximately 5 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 133.6, b = 149.9, c = 255.1 A, and space group P2(1), with unit-cell parameters a = 175.1, b = 162.6, c = 242.8 A, beta = 90.1 degrees , respectively. For unlabelled HRSVN, only crystals belonging to space group P2(1) were obtained that diffracted to 3.6 A. A self-rotation function using data from the orthorhombic crystal form confirmed the presence of tenfold noncrystallographic symmetry, which is in agreement with a reported electron-microscopic reconstruction of HRSVN. Monomeric HRSVN generated by N-terminal truncation was designed to assist in structure determination by reducing the size of the asymmetric unit. Whilst such HRSVN mutants were monomeric in solution and crystallized in a different space group, the size of the asymmetric unit was not reduced. |
| first_indexed | 2024-03-06T22:22:55Z |
| format | Journal article |
| id | oxford-uuid:55ae7ab8-77b2-4710-9df9-f30afe71fee3 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:22:55Z |
| publishDate | 2008 |
| record_format | dspace |
| spelling | oxford-uuid:55ae7ab8-77b2-4710-9df9-f30afe71fee32022-03-26T16:45:29ZCrystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:55ae7ab8-77b2-4710-9df9-f30afe71fee3EnglishSymplectic Elements at Oxford2008El Omari, KScott, KDhaliwal, BRen, JAbrescia, NBudworth, JLockyer, MPowell, KHawkins, AStammers, DHuman respiratory syncytial virus (HRSV) has a nonsegmented negative-stranded RNA genome which is encapsidated by the HRSV nucleocapsid protein (HRSVN) that is essential for viral replication. HRSV is a common cause of respiratory infection in infants, yet no effective antiviral drugs to combat it are available. Recent data from an experimental anti-HRSV compound, RSV-604, indicate that HRSVN could be the target site for drug action. Here, the expression, purification and preliminary data collection of decameric HRSVN as well as monomeric N-terminally truncated HRSVN mutants are reported. Two different crystal forms of full-length selenomethionine-labelled HRSVN were obtained that diffracted to 3.6 and approximately 5 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 133.6, b = 149.9, c = 255.1 A, and space group P2(1), with unit-cell parameters a = 175.1, b = 162.6, c = 242.8 A, beta = 90.1 degrees , respectively. For unlabelled HRSVN, only crystals belonging to space group P2(1) were obtained that diffracted to 3.6 A. A self-rotation function using data from the orthorhombic crystal form confirmed the presence of tenfold noncrystallographic symmetry, which is in agreement with a reported electron-microscopic reconstruction of HRSVN. Monomeric HRSVN generated by N-terminal truncation was designed to assist in structure determination by reducing the size of the asymmetric unit. Whilst such HRSVN mutants were monomeric in solution and crystallized in a different space group, the size of the asymmetric unit was not reduced. |
| spellingShingle | El Omari, K Scott, K Dhaliwal, B Ren, J Abrescia, N Budworth, J Lockyer, M Powell, K Hawkins, A Stammers, D Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein. |
| title | Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein. |
| title_full | Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein. |
| title_fullStr | Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein. |
| title_full_unstemmed | Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein. |
| title_short | Crystallization and preliminary X-ray analysis of the human respiratory syncytial virus nucleocapsid protein. |
| title_sort | crystallization and preliminary x ray analysis of the human respiratory syncytial virus nucleocapsid protein |
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