Description
General description
Shutoff protein (UniProt: P24933; also known as 100 kDa protein, (p100K), 100K-chaperone protein, L4-100K, Shutoff protein 100K) is encoded by the L4 gene (Gene ID: 2653005) in human Adenovirus 5. Adenovirus 5 belongs to the Adenoviridae family of non-enveloped viruses that are characterized by linear, double-stranded DNA genomes. Their capsid is an icosahedron that contains at least 12 different viral proteins. Adenovirus 5 is commonly used for gene transfer experiments due to its ability to infect a wide group of different cell types. It can harbor large genes in its genome incorporated via homologous recombination techniques. Adenovirus assembly protein L4-100K is a multifunctional protein critical for the adenovirus replication cycle. It selectively that inhibits host translation while promoting late viral translation by ribosome shunting. Also, assists in the nuclear export of late viral mRNAs, ensuring their translation in the cytoplasm. L4-100K exists as a monomer and interacts with hexon protein. This interaction allows chaperoning and trimerization of hexon proteins, aiding in their proper folding and assembly into the virion capsid. It contains a short binding region (aa 280-345) that is essential for binding to the tripartite leader sequence of viral late mRNAs and recruits host eIF4G. L4-100K is expressed in the late phase of viral replicative cycle and its expression is tightly regulated by the viral transcriptional machinery and coincides with the synthesis of other late viral proteins necessary for virion assembly. L4-100K protein is predominantly localized in the cytoplasm of infected cells and it shuttles between the nucleus and cytoplasm. Its involvement in the nuclear export of viral mRNAs, enhancement of translation, inhibition of host protein synthesis, and chaperoning of hexon proteins underscores its importance in viral replication and pathogenicity. Studies demonstrate that L4-100K is a GrB substrate that prevents cytotoxic lymphocyte granule-induced apoptosis in infected target cells by potently inhibiting GrB. 100K is the first viral protein described that exclusively targets the GrB pathway and it represents a novel class of viral protease inhibitor, in which an essential, multifunctional viral protein, which is vulnerable to specific proteolysis by GrB, expresses inhibitory function against that protease. (Ref.: Andrade, F., et al. (2001). Immunity 14 (6); 751-761; Young, A-M., et al. (2012). Mol. Ther. 20(9); 1676-1688; Yan, J., et al. (2016). Scientific Report 3(6); 22464).
Immunogen
His-tagged, recombinant fragment of human Adenovirus 5 corresponding to 446 amino acids from the C-terminal half.
Application
Quality Control Testing
Evaluated by Western Blotting in Hela cells infected with human Adenovirus 5.
Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected L4-100K (Shutoff protein) in lysate from HeLa cells infected with human Adenovirus 5, but not in lysate from uninfected HeLa cells.
Tested Applications
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected L4-100K protein of Adenovirus 5 in lysate from HeLa whole cell extract infected with HAdV-C5-WT, but not in lysate from uninfected HeLa whole cell extract (Courtesy of Dr. Patrick Hearing, Stony Brook University).
Immunoprecipitation Analysis: A representative lot immunoprecipitated Adenovirus 5 L4-100K in Immunoprecipitation (Yan, J. et al. (2016) Scientific Report 3(6); 22464).
ELISA: A representative lot detected Adenovirus 5 L4-100K in ELISA applications (Yan, J. et al. (2016) Scientific Report 3(6); 22464).
Western Blotting Analysis: A representative lot detected Adenovirus 5 L4-100K in Western Blotting applications (Young, A-M. et al. (2012) Mol. Ther. 20(9); 1676-1688).
Immunofluorescence Analysis: A representative lot detected Adenovirus 5 L4-100K in Immunofluorescence applications (Young, A-M. et al. (2012) Mol. Ther. 20(9); 1676-1688; Yan, J. et al. (2016) Scientific Report 3(6); 22464).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
This rabbit polyclonal antibody specifically detects L4-100K (Shutoff protein) of human Adenovirus 5. It targets an epitope within 446 amino acids from the C-terminal half.
Physical form
Rabbit polyclonal antiserum without azide.
Preparation Note
Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
