Synthesis and glycosylation in vitro of glycoprotein of vesicular stomatitis virus.
Synthesis and glycosylation in vitro of glycoprotein of vesicular stomatitis virus.
复制标题
水疱性口炎病毒糖蛋白的体外合成和糖基化。
DOI：
10.1073/pnas.74.4.1516
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发表时间：
1977
影响因子：
11.1
通讯作者：
H. Ghosh
中科院分区：
文献类型：
--
作者：
F. Toneguzzo;H. Ghosh
从水泡性口炎病毒感染的L细胞的核糖核蛋白颗粒的耦合到从未感染的HeLa细胞获得的预孵育的核糖体系统允许合成两种蛋白质。G1（分子量63，000）和G2（分子量67，000），以及除刺突蛋白G（分子量69，000）之外的水疱性口炎病毒的所有其他蛋白。胰蛋白酶肽的分析表明，G1，G2和G具有相同的肽序列。G2的合成需要膜的存在;只有G1在没有任何膜的情况下合成。用刀豆球蛋白A-Sepharose柱亲和层析证明G2是一种糖蛋白，而G1不是。通过神经氨酸酶从G去除唾液酸残基导致产物具有与G2相同的迁移率。然而，用神经氨酸酶（EC 3.2.1.18）、β-半乳糖苷酶（EC 3.2.1.23）和β-N-乙酰氨基葡糖苷酶（EC 3.2.1.30）的混合物消化G2或G产生分子量为65，000的蛋白质。这些数据表明，G2是去唾液酸化的G，并通过G1的糖基化形成，G1是G的未糖基化的多肽骨架。
Coupling of ribonucleoprotein particles from L cells infected with vesicular stomatitis virus to a pre-incubated ribosomal system obtained from uninfected HeLa cells allowed synthesis of two proteins. G1 (molecular weight 63,000) and G2 (molecular weight 67,000), and all other proteins of vesicular stomatitis virus except the spike protein G (molecular weight 69,000). Analyses of the tryptic peptides showed that G1, G2, and G had identical peptide sequences. The synthesis of G2 required the presence of membranes; only G1 was synthesized in the absence of any membranes. G2 but not G1 was shown to be a glycoprotein by affinity chromatography on a concanavalin A-Sepharose column. Removal of sialic acid residues from G by neuraminidase resulted in a product having an identical mobility to G2. Digestion of G2 or G with a mixture of neuraminidase (EC 3.2.1.18), beta-galactosidase (EC 3.2.1.23), and beta-N-acetylglucosaminidase (EC 3.2.1.30), however, produced a protein of molecular weight 65,000. These data suggest that G2 is the desialated G and is formed by glycosylation of G1, which is the unglycosylated polypeptide backbone of G.