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仙台病毒HN和F糖蛋白作为HTC细胞中质膜蛋白分解代谢的探针。融合性重组仙台病毒包膜的研究。

Sendai-viral HN and F glycoproteins as probes of plasma-membrane protein catabolism in HTC cells. Studies with fusogenic reconstituted Sendai-viral envelopes.

作者信息

Earl R T, Billett E E, Hunneyball I M, Mayer R J

出版信息

Biochem J. 1987 Feb 1;241(3):801-7. doi: 10.1042/bj2410801.

Abstract

Reconstituted Sendai-viral envelopes (RSVE) were produced by the method of Vainstein, Hershkovitz, Israel & Loyter [(1984) Biochim. Biophys. Acta 773, 181-188]. RSVE are fusogenic unilamellar vesicles containing two transmembrane glycoproteins: the HN (haemagglutinin-neuraminidase) protein and the F (fusion) factor. The fate of the viral proteins after fusion-mediated transplantation of RSVE into hepatoma (HTC) cell plasma membranes was studied to probe plasma-membrane protein degradation. Both protein species are degraded at similar, relatively slow, rates (t1/2 = 67 h) in HTC cells fused with RSVE in suspension. Even slower degradation rates for HN and F proteins (t1/2 = 93 h) were measured when RSVE were fused with HTC cells in monolayer. Lysosomal degradation of the transplanted viral proteins is strongly implicated by the finding that degradation of HN and F proteins is sensitive to inhibition by 10 mM-NH4Cl (81%) and by 50 micrograms of leupeptin/ml (70%).

摘要

重组仙台病毒包膜(RSVE)采用Vainstein、Hershkovitz、Israel和Loyter的方法制备[(1984年)《生物化学与生物物理学报》773卷,181 - 188页]。RSVE是含有两种跨膜糖蛋白的融合性单层囊泡:HN(血凝素 - 神经氨酸酶)蛋白和F(融合)因子。为了探究质膜蛋白降解情况,研究了RSVE通过融合介导移植到肝癌(HTC)细胞质膜后病毒蛋白的命运。在悬浮状态下与RSVE融合的HTC细胞中,这两种蛋白以相似的相对较慢速率(半衰期 = 67小时)降解。当RSVE与单层HTC细胞融合时,测得HN和F蛋白的降解速率更慢(半衰期 = 93小时)。发现HN和F蛋白的降解对10 mM - NH₄Cl(81%)和50微克亮抑酶肽/毫升(70%)的抑制敏感,这有力地表明移植的病毒蛋白是通过溶酶体降解的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b75/1147633/1f2a2adda661/biochemj00262-0174-a.jpg

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