Role of cysteines in infectious bronchitis virus envelope protein oligomerization.
Coronavirus envelope E protein plays an important role in the virus assembly and morphogenesis. Despite of little sequence homology among the coronavirus E protein family, two or three cysteines located near the C-terminal side of the transmembrane domain are conserved in all E proteins. Infectious...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2010
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| Online Access: | http://hdl.handle.net/10356/39619 |
| _version_ | 1826116170143498240 |
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| author | Huang, Lu. |
| author2 | Jaume Torres |
| author_facet | Jaume Torres Huang, Lu. |
| author_sort | Huang, Lu. |
| collection | NTU |
| description | Coronavirus envelope E protein plays an important role in the virus assembly and morphogenesis. Despite of little sequence homology among the coronavirus E protein family, two or three cysteines located near the C-terminal side of the transmembrane domain are conserved in all E proteins. Infectious bronchitis virus (IBV) has one of the longest coronavirus E proteins with108 amino acid residues and a molecular weight of 12.3 kD. It is also distinctive by having only two conserved cysteines while most other coronavirus E proteins have three. In this study, to facilitate the expression and purification process, IBV E has been fused with a modified β-barrel platform protein. The fusion protein was expressed in E. coli, recovered from inclusion bodies, cleaved by CNBr and IBV E was purified by HPLC. It is the first time to express and purify IBV E and analyze its secondary structure and oligomerization. Gel electrophoresis revealed that IBV E forms mostly monomer in SDS and pentamer in milder detergent PFO. AUC analysis of IBV E in C14 betaine revealed good fitting for monomer-pentamer model. Analysis using ATR-FTIR revealed strikingly high percentage of residues protected from H/D exchange, providing clue for the IBV E membrane topology. |
| first_indexed | 2024-10-01T04:06:58Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/39619 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:06:58Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | ntu-10356/396192023-02-28T18:00:07Z Role of cysteines in infectious bronchitis virus envelope protein oligomerization. Huang, Lu. Jaume Torres School of Biological Sciences DRNTU::Science Coronavirus envelope E protein plays an important role in the virus assembly and morphogenesis. Despite of little sequence homology among the coronavirus E protein family, two or three cysteines located near the C-terminal side of the transmembrane domain are conserved in all E proteins. Infectious bronchitis virus (IBV) has one of the longest coronavirus E proteins with108 amino acid residues and a molecular weight of 12.3 kD. It is also distinctive by having only two conserved cysteines while most other coronavirus E proteins have three. In this study, to facilitate the expression and purification process, IBV E has been fused with a modified β-barrel platform protein. The fusion protein was expressed in E. coli, recovered from inclusion bodies, cleaved by CNBr and IBV E was purified by HPLC. It is the first time to express and purify IBV E and analyze its secondary structure and oligomerization. Gel electrophoresis revealed that IBV E forms mostly monomer in SDS and pentamer in milder detergent PFO. AUC analysis of IBV E in C14 betaine revealed good fitting for monomer-pentamer model. Analysis using ATR-FTIR revealed strikingly high percentage of residues protected from H/D exchange, providing clue for the IBV E membrane topology. Bachelor of Science in Biological Sciences 2010-06-01T08:51:05Z 2010-06-01T08:51:05Z 2010 2010 Final Year Project (FYP) http://hdl.handle.net/10356/39619 en Nanyang Technological University 35 p. application/pdf |
| spellingShingle | DRNTU::Science Huang, Lu. Role of cysteines in infectious bronchitis virus envelope protein oligomerization. |
| title | Role of cysteines in infectious bronchitis virus envelope protein oligomerization. |
| title_full | Role of cysteines in infectious bronchitis virus envelope protein oligomerization. |
| title_fullStr | Role of cysteines in infectious bronchitis virus envelope protein oligomerization. |
| title_full_unstemmed | Role of cysteines in infectious bronchitis virus envelope protein oligomerization. |
| title_short | Role of cysteines in infectious bronchitis virus envelope protein oligomerization. |
| title_sort | role of cysteines in infectious bronchitis virus envelope protein oligomerization |
| topic | DRNTU::Science |
| url | http://hdl.handle.net/10356/39619 |
| work_keys_str_mv | AT huanglu roleofcysteinesininfectiousbronchitisvirusenvelopeproteinoligomerization |