Coronavirus IBV: partial amino terminal sequencing of spike polypeptide S2 identifies the sequence Arg-Arg-Phe-Arg-Arg at the cleavage site of the spike precursor propolypeptide of IBV strains Beaudette and M41.
Coronavirus IBV: partial amino terminal sequencing of spike polypeptide S2 identifies the sequence Arg-Arg-Phe-Arg-Arg at the cleavage site of the spike precursor propolypeptide of IBV strains Beaudette and M41.
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DOI：
10.1016/0168-1702(86)90037-7
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发表时间：
1986-03
期刊：
影响因子：
5
通讯作者：
Brown TD
中科院分区：
文献类型：
--
作者：
Cavanagh D;Davis PJ;Pappin DJ;Binns MM;Boursnell ME;Brown TD
禽传染性支气管炎冠状病毒的刺突蛋白包含由前糖多肽So裂解衍生的两种糖多肽S1和S2，最近已确定了Beaudette菌株的核苷酸序列（Binns M.M.等人，1985，J. Gen. Virol. 66, 719-726）。 So内的两个糖多肽的顺序是氨基末端(N)-S1-S2-羧基末端(C)。为了定位S2的N末端，我们对来自用[3H]丝氨酸标记的IBV-Beaudette和用[3H]缬氨酸、亮氨酸和异亮氨酸标记的相关菌株IBV-M41的S2进行了部分氨基酸测序。鉴定出的残基及其相对于 S2 N 末端的位置是：丝氨酸，13；缬氨酸, 6, 12;亮氨酸，前 20 个残基中没有；异亮氨酸, 2, 19。这些结果鉴定出 IBV-Beaudette 的 S2 的 N 末端为丝氨酸，距离 S1 的 N 末端有 520 个残基，不包括信号序列。紧邻残基520 So的N端侧具有序列Arg-Arg-Phe-Arg-Arg；类似的基本连接肽是其他几种病毒刺突糖蛋白的特征。推断对于IBV-Beaudette，S1包含519个残基(Mr 57.0K)或514个残基(56.2K)，如果连接肽要通过体内羧肽酶样活性去除，而S2具有625个残基(69.2K)。 IBV-M41 So 基因切割区的核苷酸测序揭示了与 IBV-Beaudette 相同的连接肽，并且 IBV-M41 S2 的前 20 个 N 端残基与 Beaudette 菌株的相同。在 Vero 细胞中生长的 IBV-Beaudette 含有一些未切割的 So；这可被 10 μg/ml 的胰蛋白酶和胰凝乳蛋白酶裂解。 IBV-M41 的 S1 的部分 N 末端分析鉴定出分别位于 N 末端位置 2 和 9 的亮氨酸和缬氨酸残基。这证实了 Binns 等人的鉴定。 (1985)，S1 的 N 末端和 IBV-Beaudette 刺突前多肽信号序列的末端。 [3H]亮氨酸标记的IBV-Beaudette膜(M)多肽的N端测序显示亮氨酸残基位于N端的8,16和22位；这些结果证实了 M.E.G. 确定的开放阅读框。布尔斯内尔等人。 (1984, Virus Res. 1, 303–313) M 的核苷酸序列。核衣壳 (n) 多肽的 N 末端似乎被封闭。
The spike protein of avian infectious bronchitis coronavirus comprises two glycopolypeptides S1 and S2 derived by cleavage of a proglycopolypeptide So, the nucleotide sequence of which has recently been determined for the Beaudette strain (Binns M.M. et al., 1985, J. Gen. Virol. 66, 719–726). The order of the two glycopolypeptides within So is aminoterminus(N)-Sl-S2-carboxyterminus(C). To locate the N-terminus of S2 we have performed partial amino acid sequencing on S2 from IBV-Beaudette labelled with [3H]serine and from the related strain IBV-M41 labelled with [3H]valine, leucine and isoleucine. The residues identified and their positions relative to the N-terminus of S2 were: serine, 13; valine, 6, 12; leucine, none in the first 20 residues; isoleucine, 2, 19. These results identified the N-terminus of S2 of IBV-Beaudette as serine, 520 residues from the N-terminus of S1, excluding the signal sequence. Immediately to the N-terminal side of residue 520 So has the sequence Arg-Arg-Phe-Arg-Arg; similar basic connecting peptides are a feature of several other virus spike glycoproteins. It was deduced that for IBV-Beaudette SI comprises 519 residues (Mr 57.0K) or 514 residues (56.2K) if the connecting peptide was to be removed by carboxypeptidase-like activity in vivo while S2 has 625 residues (69.2K). Nucleotide sequencing of the cleavage region of the So gene of IBV-M41 revealed the same connecting peptide as IBV-Beaudette and that the first 20 N-terminal residues of S2 of IBV-M41 were identical to those of the Beaudette strain. IBV-Beaudette grown in Vero cells had some uncleaved So; this was cleavable by 10 μg/ml of trypsin and of chymotrypsin. Partial N-terminal analysis of S1 from IBV-M41 identified leucine and valine residues at positions 2 and 9 respectively from the N-terminus. This confirms the identification made by Binns et al. (1985), of the N-terminus of S1 and the end of the signal sequence of the IBV-Beaudette spike propolypeptide. N-terminal sequencing of [3H]leucine-labelled IBV-Beaudette membrane (M) polypeptide showed leucine residues at positions 8,16 and 22 from the N-terminus; these results confirm the open reading frame identified by M.E.G. Boursnell et al. (1984, Virus Res. 1, 303–313) in the nucleotide sequence of M. The N-terminus of the nucleocapsid (n) polypeptide appeared to be blocked.
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DOI：
10.1073/pnas.75.6.2737
发表时间：
1978-01-01
影响因子：
11.1
作者：
GETHING, MJ;WHITE, JM;WATERFIELD, MD
通讯作者：
WATERFIELD, MD
影响因子：
4.1
作者：
PAPPIN, DJC;FINDLAY, JBC
通讯作者：
FINDLAY, JBC
影响因子：
64.8
作者：
PORTER, AG;BARBER, C;EMTAGE, JS
通讯作者：
EMTAGE, JS
影响因子：
3.7
作者：
STURMAN, LS;HOLMES, KV
通讯作者：
HOLMES, KV
影响因子：
3.8
作者：
BINNS, MM;BOURSNELL, MEG;BROWN, TDK
通讯作者：
BROWN, TDK