Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes
Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2007.
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| Format: | Thesis |
| Language: | eng |
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Massachusetts Institute of Technology
2008
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| Online Access: | http://hdl.handle.net/1721.1/40471 |
| _version_ | 1811080562281021440 |
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| author | Rosenbaum, Lara Elise |
| author2 | Kimberly Hamad-Schifferli. |
| author_facet | Kimberly Hamad-Schifferli. Rosenbaum, Lara Elise |
| author_sort | Rosenbaum, Lara Elise |
| collection | MIT |
| description | Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2007. |
| first_indexed | 2024-09-23T11:33:38Z |
| format | Thesis |
| id | mit-1721.1/40471 |
| institution | Massachusetts Institute of Technology |
| language | eng |
| last_indexed | 2024-09-23T11:33:38Z |
| publishDate | 2008 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/404712019-04-11T03:09:53Z Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes Rosenbaum, Lara Elise Kimberly Hamad-Schifferli. Massachusetts Institute of Technology. Dept. of Mechanical Engineering. Massachusetts Institute of Technology. Dept. of Mechanical Engineering. Mechanical Engineering. Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2007. Includes bibliographical references (leaf 20). Maintaining protein function at the biological-inorganic interface is a critical challenge for bionanotechnology. Specifically, nanoparticle-protein conjugates must be designed to interact with binding partners with biologically-relevant thermodynamics. Towards developing a nanoparticle-tagging system that minimizes interference with normal protein function, here we design and begin development of an assay to assess complex formation between nanoparticle-immobilized proteins and soluble binding partners. Two chaperone proteins, importin-a and importin-3 mediate classical nuclear transport, an essential and highly conserved example of protein complex formation in eukaryotic cells. Together, these two proteins form a chaperone complex that recognizes a nuclear localization signal (NLS), which is a short peptide sequence. Here, we synthesize and purify a fluorescently-labeled importin-a and a positive control for complex formation, which consists of bovine albumin serum (BSA) covalently conjugated to a fluorophore and NLS. Using these two fluorescent molecules, we can perform Forster Resonance Energy Transfer (FRET) experiments to study the kinetics and thermodynamics of these protein interactions. The development of this system will be used in future tests with the NLS-conjugated fluorescent gold nanoparticles. by Lara Elise Rosenbaum. S.B. 2008-02-27T22:29:08Z 2008-02-27T22:29:08Z 2007 2007 Thesis http://hdl.handle.net/1721.1/40471 191748948 eng M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 24 leaves application/pdf Massachusetts Institute of Technology |
| spellingShingle | Mechanical Engineering. Rosenbaum, Lara Elise Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes |
| title | Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes |
| title_full | Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes |
| title_fullStr | Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes |
| title_full_unstemmed | Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes |
| title_short | Design of an in vitro assay to optimize assembly of nanoparticle-tagged nuclear import complexes |
| title_sort | design of an in vitro assay to optimize assembly of nanoparticle tagged nuclear import complexes |
| topic | Mechanical Engineering. |
| url | http://hdl.handle.net/1721.1/40471 |
| work_keys_str_mv | AT rosenbaumlaraelise designofaninvitroassaytooptimizeassemblyofnanoparticletaggednuclearimportcomplexes |