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Purification of the putative coxsackievirus B receptor from HeLa cells
S D Carson1, N N Chapman, S M Tracy
1Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198, USA. 73632,3623@compuserve.com
Abstract:
We have identified a protein expressed by human and murine cells susceptible to coxsackievirus B3 (CVB3) infection and purified it from HeLa cells. This protein of approximately 45,000 Mr is expressed by HeLa cells and mouse fetal heart fibroblasts (susceptible to infection), and not by C3H murine fibroblasts or the human RD cell line (resistant). The protein was isolated from Triton X-100- deoxycholate lysates of HeLa cells by chromatography on concanavalin A-Sepharose, Affi-gel Blue, Phenyl Sepharose, and PBE94. The CVB3-binding fraction from PBE94 was blotted from SDS-polyacrylamide gel onto PVDF membrane for amino acid sequencing. Approximately 2 pmoles of CVB3-binding protein provided assignments for 26 consecutive residues: LSITTPEEMIEKAKGETAYLPXKFTL. This sequence corresponds neither to decay accelerating factor nor to nucleolin, both of which have previously been identified as CVB3-binding proteins, but does match two entries in GenBank. These data show that we have purified a novel CVB3-binding protein, the characteristics of which suggest the CVB group receptor has been purified. Identification of 26 amino acid residues in the protein and corresponding GenBank enteries will accelerate study of CVB tropism and the diseases caused by these viruses.
Insights
Researchers purified a novel coxsackievirus B3 (CVB3) binding protein from human cells. This protein
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Coxsackievirus B3 (CVB3) causes various human diseases.
- Identifying cellular receptors for viruses is crucial for understanding viral tropism and developing antivirals.
Purpose of the Study:
- To identify and purify the cellular receptor for coxsackievirus B3 (CVB3).
Main Methods:
- Protein purification from HeLa cell lysates using multiple chromatography techniques.
- Amino acid sequencing of the purified CVB3-binding protein.
- Bioinformatic analysis of the protein sequence against GenBank databases.
Main Results:
- A novel protein of approximately 45,000 Mr was purified from CVB3-susceptible cells.
- The purified protein bound CVB3.
- Amino acid sequencing yielded 26 residues, matching two GenBank entries but not previously identified CVB3-binding proteins.
- The protein is expressed in susceptible cells but not in resistant cell lines.
Conclusions:
- A novel CVB3-binding protein, likely the CVB group receptor, has been purified.
- The identified amino acid sequence will facilitate further studies on CVB tropism and pathogenesis.