Enterovirus 71 binding to PSGL-1 on leukocytes: VP1-145 acts as a molecular switch to control receptor interaction.
Enterovirus 71 binding to PSGL-1 on leukocytes: VP1-145 acts as a molecular switch to control receptor interaction.
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DOI：
10.1371/journal.ppat.1003511
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发表时间：
2013
期刊：
影响因子：
6.7
通讯作者：
Shimizu H
中科院分区：
文献类型：
--
作者：
Nishimura Y;Lee H;Hafenstein S;Kataoka C;Wakita T;Bergelson JM;Shimizu H
肠道病毒71型（EV71）的一些毒株通过与一种特异性受体分子P-选择素糖蛋白配体-1（PSGL-1）结合来感染白细胞。我们发现衣壳蛋白VP1内的一个氨基酸残基决定了EV71是否与PSGL-1结合。对代表性EV71毒株的衣壳序列的检查显示，PSGL-1结合病毒在衣壳蛋白VP1内的残基145处具有G或Q（VP1 - 145 G或Q），而PSGL-1非结合病毒具有VP1 - 145 E。使用定点突变，我们发现当VP1 - 145G/Q被E取代时，PSGL-1结合菌株失去了结合能力;相反，当VP1 - 145E被G或Q取代时，非结合菌株获得了结合PSGL-1的能力。在VP1 - 145处具有G/Q的病毒有效地感染白细胞细胞系Jurkat T细胞，而在该位置处具有E的病毒则没有。我们先前报道，EV71结合到PSGL-1的N-末端区域，并且该结合依赖于该区域内的硫酸化酪氨酸残基。我们推测结合依赖于病毒衣壳中带负电荷的硫酸基团和带正电荷的碱性残基之间的相互作用。病毒表面上的VP1 - 145与VP1 - 242和VP1 - 244处的保守赖氨酸残基非常接近。比较最近发表的在VP1 - 145处具有Q或E的EV71分离物的晶体结构，揭示VP1 - 145控制VP1 - 244的赖氨酸侧链的取向：对于VP1 - 145 Q，赖氨酸侧链朝向外，但对于VP1 - 145 E，赖氨酸侧链转向病毒表面。VP1 - 244的突变消除了病毒与PSGL-1的结合，VP1 - 242的突变大大降低了结合。我们提出，保守的赖氨酸残基的病毒表面负责与硫酸化的酪氨酸残基在PSGL-1的N-末端的相互作用，和VP1 - 145作为一个开关，控制PSGL-1的结合，通过调节暴露的VP1 - 244 K。肠道病毒71型（EV71）通常引起儿童轻度发热性疾病（手足口病），但有些患者会出现严重的神经系统疾病和死亡。最近在亚太地区爆发的疫情已造成数千人死亡，使EV71成为一个重大的公共卫生问题。一些EV71毒株与P-选择素糖蛋白配体-1（PSGL-1）结合并感染免疫细胞，但其他毒株不结合。我们先前发现EV71结合PSGL-1的N-末端，并且该结合依赖于N-末端的酪氨酸硫酸化，但是控制与PSGL-1相互作用的病毒因子尚未被鉴定。在我们目前的工作中，我们提出的证据表明，一个单一的氨基酸，残基145的病毒衣壳蛋白（VP1 - 145），决定是否结合病毒PSGL-1，它的功能，通过影响附近的赖氨酸残基（VP1 - 244）的病毒表面上的方向。我们认为，VP1 - 145通过改变VP1 - 244的带正电荷的赖氨酸侧链与PSGL-1的带负电荷的硫酸化N-末端的可及性来控制病毒嗜性。我们的研究结果揭示了病毒受体的相互作用，以及EV71对表达PSGL-1的白细胞的嗜性。
Some strains of enterovirus 71 (EV71), but not others, infect leukocytes by binding to a specific receptor molecule: the P-selectin glycoprotein ligand-1 (PSGL-1). We find that a single amino acid residue within the capsid protein VP1 determines whether EV71 binds to PSGL-1. Examination of capsid sequences of representative EV71 strains revealed that the PSGL-1-binding viruses had either a G or a Q at residue 145 within the capsid protein VP1 (VP1-145G or Q), whereas PSGL-1-nonbinding viruses had VP1-145E. Using site-directed mutagenesis we found that PSGL-1-binding strains lost their capacity to bind when VP1-145G/Q was replaced by E; conversely, nonbinding strains gained the capacity to bind PSGL-1 when VP1-145E was replaced with either G or Q. Viruses with G/Q at VP1-145 productively infected a leukocyte cell line, Jurkat T-cells, whereas viruses with E at this position did not. We previously reported that EV71 binds to the N-terminal region of PSGL-1, and that binding depends on sulfated tyrosine residues within this region. We speculated that binding depends on interaction between negatively charged sulfate groups and positively charged basic residues in the virus capsid. VP1-145 on the virus surface is in close proximity to conserved lysine residues at VP1-242 and VP1-244. Comparison of recently published crystal structures of EV71 isolates with either Q or E at VP1-145 revealed that VP1-145 controls the orientation of the lysine side-chain of VP1-244: with VP1-145Q the lysine side chain faces outward, but with VP1-145E, the lysine side chain is turned toward the virus surface. Mutation of VP1-244 abolished virus binding to PSGL-1, and mutation of VP1-242 greatly reduced binding. We propose that conserved lysine residues on the virus surface are responsible for interaction with sulfated tyrosine residues at the PSGL-1 N-terminus, and that VP1-145 acts as a switch, controlling PSGL-1 binding by modulating the exposure of VP1-244K. Enterovirus 71 (EV71) commonly causes mild febrile illness in children (hand, foot, and mouth disease), but some patients suffer severe neurologic disease and death. Recent outbreaks in the Asia-Pacific region have caused thousands of deaths, making EV71 a major public health concern. Some EV71 strains bind to P-selectin glycoprotein ligand-1 (PSGL-1) and infect immune cells, but others do not. We previously found that EV71 binds the PSGL-1 N-terminus, and that binding depends on tyrosine sulfation of the N-terminus, but the viral factors that control interaction with PSGL-1 have not been identified. In our present work we present evidence that a single amino acid, residue 145 of the viral capsid protein (VP1-145), determines whether a virus binds or does not bind PSGL-1, and that it functions by influencing the orientation of a nearby lysine residue (VP1-244) on the virus surface. We propose that VP1-145 controls virus tropism by changing the accessibility of the positively-charged lysine side chain of VP1-244 to the negatively charged, sulfated N-terminus of PSGL-1. Our results shed new light on virus-receptor interaction, and EV71 tropism for PSGL-1-expressing leukocytes.
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