Identification of amino acids involved in the sialidase activity of the mumps virus hemagglutinin-neuraminadase protein.
Identification of amino acids involved in the sialidase activity of the mumps virus hemagglutinin-neuraminadase protein.
复制标题
鉴定参与腮腺炎病毒血凝素神经氨酸酶蛋白唾液酸酶活性的氨基酸。
DOI：
10.1016/0042-6822(88)90072-4
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发表时间：
1988
期刊：
影响因子：
3.7
通讯作者：
Aronowski,J
中科院分区：
文献类型：
--
作者：
Waxham,MN;Aronowski,J
我们之前描述了在竞争性唾液酸酶抑制剂2-脱氧-2,3-脱氢- n -乙酰神经氨酸（DANA）的选择压力下生长获得的腮腺炎病毒O'Take株唾液酸酶缺陷变体。在本报告中，我们描述了使用相同的选择方案生产腮腺炎病毒RW株唾液酸酶缺陷变体。RW变体RW(DANA)v1的生物活性与O'Take-(DANA)v1描述的生物活性相同，包括缺乏可检测的唾液酸酶活性，不变的血凝活性，以及在感染细胞单层中细胞间融合的表达。利用二维色氨酸肽图谱和单克隆抗体结合试验对每种病毒的结构蛋白进行分析，结果表明，血凝素-神经氨酸酶（HN）蛋白发生了有限的变化，并且只有HN蛋白发生了改变。测定了RW(DANA)v1 HN的全核苷酸序列，并与RW亲本的HN序列进行了比较。两个核苷酸差异解释了两个非保守氨基酸差异；从RW到RW(DANA)v1，一个IIe到一个Thr氨基酸181和一个GIn到一个Lys氨基酸261。通过与其他几种副粘病毒的数据进行比较，我们初步确定氨基酸181是腮腺炎病毒唾液酸酶活性位点的关键残基。
We previously described sialidase-deficient variants of the O'Take strain of mumps virus obtained by growth under the selective pressure of the competitive sialidase inhibitor 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (DANA). In this report, we describe the production of a sialidase-deficient variant of the RW strain of mumps virus using an identical selection protocol. The biologic activities of the RW variant, RW(DANA)v1, were identical to those described for O'Take-(DANA)v1 and included a lack of detectable sialidase activity, unchanged hemagglutination activity, and expression of cell-to-cell fusion in infected cell monolayers. Analysis of the structural proteins of each virus by both two-dimensional tryptic peptide mapping and monoclonal antibody binding assays suggested that limited changes occurred in the hemagglutinin-neuraminidase (HN) proteins and that only the HN proteins were altered. The complete nucleotide sequence of the RW(DANA)v1 HN was determined and compared to the HN sequence of the RW parent. Two nucleotide differences accounting for two nonconservative amino acid differences were noted; an IIe to a Thr at amino acid 181 and a GIn to Lys at amino acid 261 from RW to RW(DANA)v1, respectively. By comparing the data presented here with those reported for several other paramyxoviruses, we tentatively identify amino acid 181 as a critical residue in the active site of the mumps virus sialidase enzyme.