Carson S D, Chapman N N, Tracy S M
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198, USA. 73632,
Biochem Biophys Res Commun. 1997 Apr 17;233(2):325-8. doi: 10.1006/bbrc.1997.6449.
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.
我们已经鉴定出一种由对柯萨奇病毒B3（CVB3）感染敏感的人和鼠细胞表达的蛋白质，并从HeLa细胞中纯化了它。这种分子量约为45,000的蛋白质由HeLa细胞和小鼠胎儿心脏成纤维细胞（易受感染）表达，而不由C3H鼠成纤维细胞或人RD细胞系（抗性）表达。通过伴刀豆球蛋白A-琼脂糖、Affi-凝胶蓝、苯基琼脂糖和PBE94色谱法从HeLa细胞的Triton X-100-脱氧胆酸盐裂解物中分离该蛋白质。将来自PBE94的CVB3结合部分从SDS-聚丙烯酰胺凝胶转移到PVDF膜上进行氨基酸测序。大约2皮摩尔的CVB3结合蛋白提供了26个连续残基的序列：LSITTPEEMIEKAKGETAYLPXKFTL。该序列既不对应于衰变加速因子，也不对应于核仁素，这两者先前都已被鉴定为CVB3结合蛋白，但确实与GenBank中的两个条目匹配。这些数据表明我们已经纯化了一种新型的CVB3结合蛋白，其特征表明CVB组受体已被纯化。鉴定该蛋白质中的26个氨基酸残基和相应的GenBank条目将加速对CVB嗜性以及由这些病毒引起的疾病的研究。
