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乙型流感病毒的多肽组成以及与纯化病毒颗粒相关的酶

Polypeptide composition of Influenza B viruses and enzymes associated with the purified virus particles.

作者信息

Oxford J S

出版信息

J Virol. 1973 Oct;12(4):827-35. doi: 10.1128/JVI.12.4.827-835.1973.

Abstract

Influenza B/LEE/40, B/Rome/1/67, B/Hong Kong/8/73, and B/Victoria/98926/70 viruses have a similar polypeptide composition as analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These viruses are composed of six or seven polypeptides, depending on whether one or two high-molecular-weight polypeptides are resolved, ranging in molecular weights from 27,000 to 90,400. Three of these polypeptides, namely the heavy and light hemagglutinin chains and the neuraminidase, have attached carbohydrate. Highly purified influenza B/LEE/40 and B/Rome/1/67 virus preparations have RNA-dependent RNA polymerase activity equivalent to the incorporation of 100 and 30 pmol, respectively, of (3)H-UMP per mg of virus protein per h at 37 C, which is demonstrated only in detergent-treated virus suspensions. However, no RNA-dependent DNA polymerase enzyme activity was detected in the two viruses although virus suspensions were "activated" by heat, alpha-chymotrypsin, and detergents. Other enzymatic activities were associated with purified preparations of influenza B virus and were attributed to minor contamination of virus with host cell enzymes. Thus, nucleoside and deoxynucleoside phosphohydrolase enzymes were active in the absence of detergents and catalyzed the release of 1,200 and 1,800 nmol of P(i) per mg of virus protein in 30 min at 37 C from ATP and dATP substrates. Thin-layer chromatography indicated that the products of the phosphohydrolase enzymes of influenza B/LEE/40 were mainly nucleoside diphosphate and monophosphate. The latter enzymes were tightly bound to influenza B/LEE/40 virus and could not be removed completely by repeated centrifugation, including centrifugation of the virus to equilibrium in density gradients of 25 to 40% (wt/vol) cesium chloride. A low degree of RNase (approximately 0.01 mug% contamination) and phosphatase (10-30 nmol of P(i) released per mg of virus protein per 30 min) activity was detected in some, but not all, influenza B/LEE/40 virus preparations.

摘要

通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析,乙型流感病毒/李/40、乙型流感病毒/罗马/1/67、乙型流感病毒/香港/8/73和乙型流感病毒/维多利亚/98926/70具有相似的多肽组成。这些病毒由六种或七种多肽组成,这取决于是否能分辨出一种或两种高分子量多肽,其分子量范围为27,000至90,400。其中三种多肽,即重链和轻链血凝素以及神经氨酸酶,带有连接的碳水化合物。高度纯化的乙型流感病毒/李/40和乙型流感病毒/罗马/1/67病毒制剂在37℃下每毫克病毒蛋白每小时具有相当于分别掺入100和30皮摩尔(3)H-UMP的RNA依赖性RNA聚合酶活性,这仅在经去污剂处理的病毒悬液中得到证实。然而,尽管病毒悬液经加热、α-胰凝乳蛋白酶和去污剂“激活”,但在这两种病毒中均未检测到RNA依赖性DNA聚合酶活性。其他酶活性与纯化的乙型流感病毒制剂相关,并归因于病毒被宿主细胞酶的轻微污染。因此,核苷和脱氧核苷磷酸水解酶在没有去污剂的情况下具有活性,并在37℃下30分钟内每毫克病毒蛋白从ATP和dATP底物催化释放1200和1800纳摩尔的无机磷酸。薄层色谱表明,乙型流感病毒/李/40的磷酸水解酶的产物主要是核苷二磷酸和单磷酸。后一种酶与乙型流感病毒/李/40病毒紧密结合,不能通过反复离心完全去除,包括将病毒在25%至40%(重量/体积)氯化铯密度梯度中离心至平衡。在一些但不是所有的乙型流感病毒/李/40病毒制剂中检测到低水平的核糖核酸酶(约0.01微克%污染)和磷酸酶(每毫克病毒蛋白每30分钟释放10 - 30纳摩尔无机磷酸)活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d0e/356700/27bd62ca72c0/jvirol00262-0171-a.jpg

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