Draft crystal structure of the vault shell at 9-A resolution.
Vaults are the largest known cytoplasmic ribonucleoprotein structures and may function in innate immunity. The vault shell self-assembles from 96 copies of major vault protein and encapsulates two other proteins and a small RNA. We crystallized rat liver vaults and several recombinant vaults, all am...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2007-11-01
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| Series: | PLoS Biology |
| Online Access: | http://europepmc.org/articles/PMC2229873?pdf=render |
| _version_ | 1818624013089374208 |
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| author | Daniel H Anderson Valerie A Kickhoefer Stuart A Sievers Leonard H Rome David Eisenberg |
| author_facet | Daniel H Anderson Valerie A Kickhoefer Stuart A Sievers Leonard H Rome David Eisenberg |
| author_sort | Daniel H Anderson |
| collection | DOAJ |
| description | Vaults are the largest known cytoplasmic ribonucleoprotein structures and may function in innate immunity. The vault shell self-assembles from 96 copies of major vault protein and encapsulates two other proteins and a small RNA. We crystallized rat liver vaults and several recombinant vaults, all among the largest non-icosahedral particles to have been crystallized. The best crystals thus far were formed from empty vaults built from a cysteine-tag construct of major vault protein (termed cpMVP vaults), diffracting to about 9-A resolution. The asymmetric unit contains a half vault of molecular mass 4.65 MDa. X-ray phasing was initiated by molecular replacement, using density from cryo-electron microscopy (cryo-EM). Phases were improved by density modification, including concentric 24- and 48-fold rotational symmetry averaging. From this, the continuous cryo-EM electron density separated into domain-like blocks. A draft atomic model of cpMVP was fit to this improved density from 15 domain models. Three domains were adapted from a nuclear magnetic resonance substructure. Nine domain models originated in ab initio tertiary structure prediction. Three C-terminal domains were built by fitting poly-alanine to the electron density. Locations of loops in this model provide sites to test vault functions and to exploit vaults as nanocapsules. |
| first_indexed | 2024-12-16T18:50:12Z |
| format | Article |
| id | doaj.art-a040732d35f74bf2bc2a5a60314182f7 |
| institution | Directory Open Access Journal |
| issn | 1544-9173 1545-7885 |
| language | English |
| last_indexed | 2024-12-16T18:50:12Z |
| publishDate | 2007-11-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj.art-a040732d35f74bf2bc2a5a60314182f72022-12-21T22:20:43ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852007-11-01511e31810.1371/journal.pbio.0050318Draft crystal structure of the vault shell at 9-A resolution.Daniel H AndersonValerie A KickhoeferStuart A SieversLeonard H RomeDavid EisenbergVaults are the largest known cytoplasmic ribonucleoprotein structures and may function in innate immunity. The vault shell self-assembles from 96 copies of major vault protein and encapsulates two other proteins and a small RNA. We crystallized rat liver vaults and several recombinant vaults, all among the largest non-icosahedral particles to have been crystallized. The best crystals thus far were formed from empty vaults built from a cysteine-tag construct of major vault protein (termed cpMVP vaults), diffracting to about 9-A resolution. The asymmetric unit contains a half vault of molecular mass 4.65 MDa. X-ray phasing was initiated by molecular replacement, using density from cryo-electron microscopy (cryo-EM). Phases were improved by density modification, including concentric 24- and 48-fold rotational symmetry averaging. From this, the continuous cryo-EM electron density separated into domain-like blocks. A draft atomic model of cpMVP was fit to this improved density from 15 domain models. Three domains were adapted from a nuclear magnetic resonance substructure. Nine domain models originated in ab initio tertiary structure prediction. Three C-terminal domains were built by fitting poly-alanine to the electron density. Locations of loops in this model provide sites to test vault functions and to exploit vaults as nanocapsules.http://europepmc.org/articles/PMC2229873?pdf=render |
| spellingShingle | Daniel H Anderson Valerie A Kickhoefer Stuart A Sievers Leonard H Rome David Eisenberg Draft crystal structure of the vault shell at 9-A resolution. PLoS Biology |
| title | Draft crystal structure of the vault shell at 9-A resolution. |
| title_full | Draft crystal structure of the vault shell at 9-A resolution. |
| title_fullStr | Draft crystal structure of the vault shell at 9-A resolution. |
| title_full_unstemmed | Draft crystal structure of the vault shell at 9-A resolution. |
| title_short | Draft crystal structure of the vault shell at 9-A resolution. |
| title_sort | draft crystal structure of the vault shell at 9 a resolution |
| url | http://europepmc.org/articles/PMC2229873?pdf=render |
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