- MeSH
- akutní myeloidní leukemie etiologie MeSH
- chytrý telefon MeSH
- dítě MeSH
- elektromagnetická pole škodlivé účinky MeSH
- elektromagnetické záření MeSH
- lidé MeSH
- magnetické pole škodlivé účinky MeSH
- matka - expozice noxám MeSH
- počítače do ruky MeSH
- radiační expozice prevence a kontrola škodlivé účinky MeSH
- zpožděný efekt prenatální expozice MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
Retroviral gag proteins, as well as fragments minimally containing the capsid (CA) and nucleocapsid (NC) subunits of Gag, are able to spontaneously assemble into virus-like particles (VLPs). This occurs in mammalian and bacterial cells as well as in in vitro systems. In every circumstance, nucleic acids are incorporated into the forming particles. Here, we took advantage of an in vitro system for the generation of non-enveloped Mason-Pfizer monkey virus (M-PMV) VLPs derived from a self-assembling CA-NC subunit of Gag. These VLPs were modified through N-terminal extension of CA-NC with short oligopeptides that, after the assembly process, were exposed on the surface of VLPs. The employed N-terminal modifications allowed specific interaction with target cells expressing prostate-specific membrane antigen. Using this system, we were able to incorporate selected siRNA into forming VLPs and deliver it into the cytosol of target cells. In comparison with other viral vectors designed for targeted transgene delivery, this M-PMV VLP system represents the lowest risk of generating virus-associated pathology, as the VLPs do not contain any viral coding sequences and are formed in a cell-free system.
- MeSH
- antigeny povrchové metabolismus MeSH
- buněčné linie MeSH
- glutamátkarboxypeptidasa II metabolismus MeSH
- lidé MeSH
- makromolekulární látky metabolismus MeSH
- malá interferující RNA metabolismus MeSH
- Masonův-Pfizerův opičí virus genetika MeSH
- přichycení viru * MeSH
- transdukce genetická * MeSH
- virům podobné částice genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This article describes basic principles of retroviral shell architecture and the approaches to its studies. Elucidation of the retrovirus particle structure, both in immature and mature forms, is essential for designing compounds that could block interactions at interfaces of viral proteins, which are critical for formation and stability of the particle. Detailed understanding of HIV structure could thus be a significant tool in the search for novel types of HIV inhibitors targeted on the assembly or maturation of this virus.
- Klíčová slova
- retrovirus, skládání retrovirové částice, struktura retrovirové částice, M-PMV, kryo-elektronová mikroskopie,
- MeSH
- elektronová mikroskopie MeSH
- genové produkty gag MeSH
- HIV infekce MeSH
- HIV MeSH
- Masonův-Pfizerův opičí virus MeSH
- molekulární sekvence - údaje MeSH
- Retroviridae fyziologie růst a vývoj MeSH
- terciární struktura proteinů MeSH
- zobrazování trojrozměrné MeSH
- Publikační typ
- práce podpořená grantem MeSH
Heterologous proteins are frequently purified by immobilized metal ion affinity chromatography (IMAC) based on their modification with a hexa-histidine affinity tag (His-tag). The terminal His-tag can, however, alter functional properties of the tagged protein. Numerous strategies for the tag removal have been developed including chemical treatment and insertion of protease target sequences in the protein sequence. Instead of using these approaches, we took an advantage of natural interaction of zinc finger domains with metal ions to purify functionally similar retroviral proteins from two different retroviruses. We found that these proteins exhibited significantly different affinities to the immobilized metal ions, despite that both contain the same type of zinc finger motif (i.e., CCHC). While zinc finger proteins may differ in biochemical properties, the multitude of IMAC platforms should allow relatively simple yet specific method for their isolation in native state.
- MeSH
- chromatografie afinitní metody MeSH
- Escherichia coli genetika MeSH
- exprese genu MeSH
- HIV-1 chemie genetika MeSH
- kovy chemie MeSH
- Masonův-Pfizerův opičí virus chemie genetika MeSH
- virové proteiny chemie genetika izolace a purifikace MeSH
- zinek analýza MeSH
- zinkové prsty MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
