Related Experiment Videos
A 70 kDa MHC class I associated protein (MAP-70) identified as a receptor molecule for Coxsackievirus A9 cell
M Triantafilou1, K Triantafilou, K M Wilson
1Department of Biological Sciences, University of Essex, Colchester, Essex, United Kingdom. mtrian@essex.ac.uk
Abstract:
One of the major categories of disease-causing micro-organisms are viruses. New studies on many different viruses have shown that virus attachment and cell entry is often a multistep process, requiring many interactions between the virus and cell surface molecules. In this study, we have attempted to identify the cell surface molecules involved in Coxsackievirus A9 (CAV-9), a common human pathogen and a member of the Picornavirus family, infectious process. GMK cells susceptible to virus infection were surfaced labeled with biotin and then solubilized in non-ionic and zwiterionic detergents. Free CAV-9 virions were used as an affinity surface, allowing the virus to bind to the solubilized receptors. The virus-receptor complexes were then immunoprecipitated by an anti CAV-9 serum and protein-A sepharose beads. SDS-PAGE and two-dimensional electrophoresis revealed the presence of integrin alpha v beta 3 molecules and a 70 kDa protein with apparent isoelectric point (pI) 5.5. The identity of the integrin alpha v beta 3 molecules was confirmed by immunoprecipitation and Western blotting; whereas the 70 kDa protein was also found to co-immunoprecipitate with MHC class I molecules in non-stringent conditions. Sequential immunoprecipitation experiments confirmed that the MHC class I associated protein (MAP-70) and the 70 kDa protein utilized by CAV-9 were identical. The role of MAP-70 in CAV-9 infectious process is discussed.
Insights
Researchers identified key cell surface molecules involved in Coxsackievirus A9 (CAV-9) infection. Integrin alpha v beta 3 and MHC class I associated protein (MAP-70) were found to be crucial for CAV-9 attachment and entry into host cells.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Viruses cause disease and their infection involves complex interactions between viral and host cell surface molecules.
- Coxsackievirus A9 (CAV-9), a Picornavirus, is a common human pathogen.
- Understanding virus-host interactions is crucial for developing antiviral strategies.
Purpose of the Study:
- To identify the specific cell surface molecules utilized by Coxsackievirus A9 (CAV-9) during its infectious process.
- To elucidate the role of these molecules in viral attachment and cell entry.
Main Methods:
- GMK cells were surface-labeled and solubilized.
- Coxsackievirus A9 (CAV-9) virions were used to affinity-purify binding receptors.
- Virus-receptor complexes were immunoprecipitated using anti-CAV-9 serum and protein-A sepharose.
- SDS-PAGE, 2D electrophoresis, immunoprecipitation, and Western blotting were employed for analysis.
Main Results:
- Integrin alpha v beta 3 and a 70 kDa protein were identified as potential CAV-9 receptors.
- The 70 kDa protein was confirmed to be MHC class I associated protein (MAP-70).
- Both integrin alpha v beta 3 and MAP-70 were found to be involved in CAV-9 infection.
Conclusions:
- Integrin alpha v beta 3 and MAP-70 are critical cell surface molecules for Coxsackievirus A9 (CAV-9) entry.
- This study provides insights into the molecular mechanisms of CAV-9 pathogenesis.
- Further research into the role of MAP-70 could lead to novel therapeutic targets.