Molecular mechanism of SCARB2-mediated attachment and uncoating of EV71

Dang, Minghao; Wang, Xiangxi; Wang, Quan; Wang, Yaxin; Jiao, Lin; Sun, Yuna; Li, Xuemei; Zhang, Liguo; Lou, Zhiyong; Wang, Junzhi; Rao, Zihe

doi:10.1007/s13238-014-0087-3

Cited by 105 publications

(112 citation statements)

References 49 publications

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“…It was recently suggested that the canyon region of EV71 mediates its interaction with SCARB2 (18). Specifically, five synthetic peptides whose sequences were located around the canyon, according to the three-dimensional structure of EV71, including VP1-2 (residues 92 to 106), VP1-6 (residues 212 to 225), VP1-8 (residues 280 to 293), VP2-2 (residues 136 to 150), and VP3-4 (residues 175 to 188), were found to bind SCARB2 in pulldown assays (18) (Fig. 2E).…”

Section: Discussionmentioning

confidence: 99%

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Single Neutralizing Monoclonal Antibodies Targeting the VP1 GH Loop of Enterovirus 71 Inhibit both Virus Attachment and Internalization during Viral Entry

Shi

et al. 2015

J Virol

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Antibodies play a critical role in immunity against enterovirus 71 (EV71). However, how EV71-specific antibodies neutralize infections remains poorly understood. Here we report the working mechanism for a group of three monoclonal antibodies (MAbs) that potently neutralize EV71. We found that these three MAbs (termed D5, H7, and C4, respectively) recognize the same conserved neutralizing epitope within the VP1 GH loop of EV71. Single MAbs in this group, exemplified by D5, could inhibit EV71 infection in cell cultures at both the pre-and postattachment stages in a cell type-independent manner. Specifically, MAb treatment resulted in the blockade of multiple steps of EV71 entry, including virus attachment, internalization, and subsequent uncoating and RNA release. Furthermore, we show that the D5 and C4 antibodies can interfere with EV71 binding to its key receptors, including heparan sulfate, SCARB2, and PSGL-1, thus providing a possible explanation for the observed multi-inhibitory function of the MAbs. Collectively, our study unravels the mechanism of neutralization by a unique group of anti-EV71 MAbs targeting the conserved VP1 GH loop. The findings should enhance our understanding of MAb-mediated immunity against enterovirus infections and accelerate the development of MAb-based anti-EV71 therapeutic drugs. IMPORTANCEEnterovirus 71 (EV71) is a major causative agent of hand, foot, and mouth disease (HFMD), which has caused significant morbidities and mortalities in young children. Neither a vaccine nor an antiviral drug is available. Neutralizing antibodies are major protective components in EV71 immunity. Here, we unraveled an unusual mechanism of EV71 neutralization by a group of three neutralizing monoclonal antibodies (MAbs). All of these MAbs bound the same conserved epitope located at the VP1 GH loop of EV71. Interestingly, mechanistic studies showed that single antibodies in this MAb group could block EV71 attachment and internalization during the viral entry process and interfere with EV71 binding to heparan sulfate, SCARB2, and PSGL-1 molecules, which are key receptors involved in different steps of EV71 entry. Our findings greatly enhance the understanding of the interplays among EV71, neutralizing antibodies, and host receptors, which in turn should facilitate the development of an MAbbased anti-EV71 therapy. E nterovirus 71 (EV71) is one of the major causative agents of hand, foot, and mouth disease (HFMD), which has been prevalent in Southeast Asia (1). Patients with severe HFMD cases manifest neurological complications, such as brainstem encephalitis and pulmonary edema, resulting in death, and such cases are often associated with EV71 infection (2). No prophylactic vaccine against EV71 infection or therapeutic drug for EV71 infection is available. Even though inactivated whole-virus-based EV71 vaccines have progressed into clinical trials (3, 4), their licensure faces serious challenges (5). Thus, the development of efficacious antiviral drugs for the treatment of EV71-infected patients ...

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Section: Discussionmentioning

confidence: 99%

“…RNA (10,18). PSGL-1 facilitates virus infection of leukocytes in a strain-specific manner (19,20); however, its exact role in viral entry in nonleukocytes remains unclear.…”

mentioning

confidence: 99%

Single Neutralizing Monoclonal Antibodies Targeting the VP1 GH Loop of Enterovirus 71 Inhibit both Virus Attachment and Internalization during Viral Entry

Shi

et al. 2015

J Virol

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“…In this study we show that alterations in helix 5 but not in helix 7 induce the fragmentation of LIMP-2. These helices are part of a helical bundle mainly formed by the three a-helices 4, 5 and 7 [2] composing the ligand binding site of LIMP-2 for interaction with GC [2] or enteroviruses [32]. It has been proposed that the helical bundle adopts two different pH-dependent conformations [33].…”

Section: Discussionmentioning

confidence: 99%

Lysosomal integral membrane protein type-2 (LIMP-2/SCARB2) is a substrate of cathepsin-F, a cysteine protease mutated in type-B-Kufs-disease

Peters

Rittger

Knabbe

et al. 2015

Biochemical and Biophysical Research Communications

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“…Our previous crystal structure of unenveloped HAV (7) showed no trace of the canyon that encircles the fivefold axes of enteroviruses and is often the site of receptor binding (8,9). Indeed, the structure provided no clues as to where T-cell Ig and mucin 1 (TIM-1), the proposed receptor (10,11), might attach.…”

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confidence: 95%

Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site

Wang

Zhu

Dang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Hepatitis A virus (HAV) infects ∼1.4 million people annually and, although there is a vaccine, there are no licensed therapeutic drugs. HAV is unusually stable (making disinfection problematic) and little is known of how it enters cells and releases its RNA. Here we report a potent HAV-specific monoclonal antibody, R10, which neutralizes HAV infection by blocking attachment to the host cell. High-resolution cryo-EM structures of HAV full and empty particles and of the complex of HAV with R10 Fab reveal the atomic details of antibody binding and point to a receptor recognition site at the pentamer interface. These results, together with our observation that the R10 Fab destabilizes the capsid, suggest the use of a receptor mimic mechanism to neutralize virus infection, providing new opportunities for therapeutic intervention.picornavirus | entry | neutralizing mechanism | receptor recognition | uncoating H epatitis A virus (HAV) is an ancient and ubiquitous pathogen found in primates and small mammals (1). It is a picornavirus with many distinctive features: in the blood it is found in an enveloped form but is shed in feces as a naked, unenveloped particle (2); it possesses a low G/C ratio in the genome sequence and a strong codon bias (3); it grows poorly in tissue culture; it possesses a 67-residue carboxyl-terminal extension of VP1 (VP1-2A or VPX), which is important for viral assembly (4); and it has a very short, nonmyristoylated VP4 (∼23 residues) (5). Because of its unusual properties, HAV remains enigmatic and occupies an evolutionary position on the periphery of picornaviruses (6, 7).Our previous crystal structure of unenveloped HAV (7) showed no trace of the canyon that encircles the fivefold axes of enteroviruses and is often the site of receptor binding (8, 9). Indeed, the structure provided no clues as to where T-cell Ig and mucin 1 (TIM-1), the proposed receptor (10, 11), might attach. In addition, the virus capsid contains no pocket factor and can withstand remarkably high temperature and low pH, indicating an uncoating mechanism (7) unlike that of enteroviruses; heating of HAV particles does not transform virions to an expanded state in vitro. Structural characterization of a HAV-TIM-1 complex might elucidate the cell-entry mechanism, but difficulty in obtaining homogenous complex preparations because of the low binding affinity of TIM-1, at least for unenveloped particles, has made this challenging. However, it is known that neutralizing antibodies (NAbs) protect against virus infection by mechanisms that include blocking attachment to the cellular receptor, overstabilizing the virus, preventing viral genome release, or physically destabilizing the virus (12, 13). Structural studies of virus complexes with such antibodies can therefore help illuminate these underlying biological functions. Here we identify a highly potent Nab, R10, which can neutralize HAV infection efficiently by blocking attachment to the host cell. The complex structure of the HAV full particle and R10 Fab suggest where the re...

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Molecular mechanism of SCARB2-mediated attachment and uncoating of EV71

Cited by 105 publications

References 49 publications

Single Neutralizing Monoclonal Antibodies Targeting the VP1 GH Loop of Enterovirus 71 Inhibit both Virus Attachment and Internalization during Viral Entry

Single Neutralizing Monoclonal Antibodies Targeting the VP1 GH Loop of Enterovirus 71 Inhibit both Virus Attachment and Internalization during Viral Entry

Lysosomal integral membrane protein type-2 (LIMP-2/SCARB2) is a substrate of cathepsin-F, a cysteine protease mutated in type-B-Kufs-disease

Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site

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