Structural definition of a neutralization-sensitive epitope on the MERS-CoV S1-NTD
Wang N, Rosen O, Wang L, Turner HL, Stevens LJ, Corbett KS, Bowman CA, Pallesen J, Shi W, Zhang Y, Leung K, Kirchdoerfer RN, Becker MM, Denison MR, Chappell JD, Ward AB, Graham BS, McLellan JS.
Cell Rep. 2019 28(13):3395-3405. Full text PDF
Middle East respiratory syndrome coronavirus (MERS-CoV) emerged into the human population in 2012 and has caused substantial morbidity and mortality. Potently neutralizing antibodies targeting the receptor-binding domain (RBD) on MERS-CoV spike (S) protein have been characterized, but much less is known about antibodies targeting non-RBD epitopes. In this manuscript, we reported the structural and functional characterization of G2, the first isolated neutralizing antibody targeting the MERS-CoV S1 N-terminal domain (S1-NTD). Structures of G2 alone, in complex with the MERS-CoV S1-NTD, and in complex with MERS-CoV S0 define a site of vulnerability that is relatively well conserved. Cell-surface binding studies, in vitro competition experiments, and neutralizing assays based on pseudovirus and authentic MERS-CoV demonstrate that G2 strongly disrupts the attachment of MERS-CoV S to its receptor, dipeptidyl peptidase-4 (DPP4), with the inhibition requiring the native trimeric S conformation. These results advance our understanding of antibody-mediated neutralization of coronaviruses and should facilitate the development of immunotherapeutics and vaccines against MERS-CoV.
The 3.1 Å cryo-electron microscopy structure of the porcine epidemic diarrhea virus spike protein in the prefusion conformation
Wrapp D & McLellan JS.
J Virol. 2019. pii: JVI.00923-19 Full text
Porcine epidemic diarrhea virus (PEDV) causes severe enteric disease in pigs, resulting in a mortality rate that exceeds 95% in neonatal piglets. PEDV is an alphacoronavirus that makes use of a large macromolecular machine called “spike” (S) to fuse the viral membrane with that of the host cell. In this manuscript, we describe the 3.1-Å cryo-EM structure of PEDV S. This structure exhibited an alternative conformation of the carbohydrate-binding attachment domain, which was not observed in the only other alphacoronavirus spike that has been described. Furthermore, careful analysis of our cryo-EM dataset revealed dissociated fusion-suppressive S1 caps that displayed their putative receptor-binding domains (RBD) in a receptor-accessible conformation. This phenomenon has never been reported for a coronavirus spike outside of the betacoronavirus genus and our findings suggest that this hinge-like RBD motion may be a conserved mechanism that is shared throughout the Coronaviridae.
Structure of the respiratory syncytial virus polymerase complex
Gilman MSA, Liu C, Fung A, Behera I, Jordan P, Rigaux P, Ysebaert N, Tcherniuk S, Sourimant J, Eléouët J-F, Sutto-Ortiz P, Decroly E, Roymans D, Jin Z, McLellan JS.
Cell 2019 179(1):193-204. Full text
Although Respiratory syncytial virus (RSV) is a leading cause of infant mortality worldwide, the only clinical intervention currently available is passive prophylaxis with the monoclonal antibody Synagis. The viral transcription and replication machinery provide an attractive target for the development of novel therapeutic agents, but structural information about this complex has remained elusive. This manuscript describes the 3.2-Å cryo-EM structure of the RSV polymerase complex, composed of the RNA-dependent RNA polymerase (L) and the tetrameric phosphoprotein (P). The structure reveals a striking tentacular arrangement of P, with each of the four monomers adopting a distinct conformation. The structure also rationalizes inhibitor escape mutants and mutations observed in live-attenuated vaccine candidates. These results provide a framework for determining the molecular underpinnings of RSV replication and transcription and should facilitate the design of effective RSV inhibitors.
Transient opening of trimeric prefusion RSV F proteins
Gilman MSA, Furmanova-Hollenstein P, Pascual G, van ‘t Wout AB, Langedijk JPM, McLellan JS.
Nat Commun. 2019 10(1):2105. Full text PDF
The respiratory syncytial virus F glycoprotein is a class I fusion protein that mediates viral entry and is the target of a number of clinical interventions currently in development for RSV. This manuscript demonstrates for the first time that the prefusion RSV F trimer transiently dissociates. The serendipitous discovery of a monoclonal antibody that favors trimer dissociation, CR9501, allowed us to determine that the prefusion RSV F trimer is in a monomer–trimer equilibrium in solution and when expressed on the surface of mammalian cells. Structural studies reveal that the prefusion F trimer samples at least two conformations, one of which reduced contacts between prefusion F monomers and might favor trimer opening. These results also have implications for RSV vaccine design, since several vaccines in clinical development are based on trimeric prefusion-stabilized F antigens.
Jones HG, Ritschel T, Pascual G, Brakenhoff JPJ, Keogh E, Furmanova-Hollenstein P, Lanckacker E, Wadia JS, Gilman MSA, Williamson RA, Roymans D, van 't Wout AB, Langedijk JP, McLellan JS.
Structural basis for recognition of the central conserved region of RSV G by neutralizing human antibodies
PLoS Pathog. 2018 14(3):e1006935.
Goodwin E, Gilman MSA, Wrapp D, Chen M, Ngwuta JO, Moin SM, Bai P, Sivasubramanian A, Connor RI, Wright PF, Graham BS, McLellan JS, Walker LM.
Infants infected with respiratory syncytial virus generate potent neutralizing antibodies that lack somatic hypermutation.
Immunity 2018; 48(2):339-349.e5.
Battles MB, Más V, Olmedillas E, Cano O, Vázquez M, Rodríguez L, Melero JA, McLellan JS.
Structure and immunogenicity of prefusion-stabilized human metapneumovirus F glycoprotein.
Nat Commun. 2017 Nov 16;8(1):1528.
Tian D, Battles MB, Moin SM, Chen M, Modjarrad K, Kumar A, Kanekiyo M, Graepel KW, Taher NM, Hotard AL, Moore ML, Zhao M, Zheng ZZ, Xia NS, McLellan JS, Graham BS.
Structural basis of respiratory syncytial virus subtype-dependent neutralization by an antibody targeting the fusion glycoprotein.
Nat Commun. 2017 Nov 30;8(1):1877.
Pallesen J, Wang N, Corbett KS, Wrapp D, Kirchdoerfer RN, Turner HL, Cottrell CA, Becker MM, Wang L, Shi W, Kong WP, Andres EL, Kettenbach AN, Denison MR, Chappell JD, Graham BS, Ward AB, McLellan JS.
Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen
PNAS 2017 Aug 29;114(35):E7348-E7357.
Rossey I, Gilman MS, Kabeche SC, Sedeyn K, Wrapp D, Kanekiyo M, Chen M, Mas V, Spitaels J, Melero JA, Graham BS, Schepens B, McLellan JS, Saelens X.
Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state.
Nat Commun. 2017 Feb 13;8:14158.
Gilman MS, Castellanos CA, Chen M, Ngwuta JO, Goodwin E, Moin SM, Mas V, Melero JA, Wright PF, Graham BS, McLellan JS, Walker LM.
Rapid profiling of RSV antibody repertoires from the memory B cells of naturally infected adult donors.
Sci Immunol. 2016 Dec 16;1(6). pii: eaaj1879.
Más V, Rodriguez L, Olmedillas E, Cano O, Palomo C, Terron MC, Luque D, Melero JA, McLellan JS.
Engineering, structure and immunogenicity of the human metapneumovirus F protein in the postfusion conformation.
PLoS Pathog. 2016 Sep 9;12(9):e1005859.
Misasi J, Gilman MS, Kanekiyo M, Gui M, Cagigi A, Mulangu S, Corti D, Ledgerwood JE, Lanzavecchia A, Cunningham J, Muyembe-Tamfun JJ, Baxa U, Graham BS, Xiang Y, Sullivan NJ, McLellan JS.
Structural and molecular basis for Ebola virus neutralization by protective human antibodies.
Science. 2016 Mar 18;351(6279):1343-6.
Kirchdoerfer RN, Cottrell CA, Wang N, Pallesen J, Yassine HM, Turner HL, Corbett KS, Graham BS, McLellan JS, Ward AB.
Pre-fusion structure of a human coronavirus spike protein.
Nature. 2016 Mar 3;531(7592):118-21.
Battles MB, Langedijk JP, Furmanova-Hollenstein P, Chaiwatpongsakorn S, Costello HM, Kwanten L, Vranckx L, Vink P, Jaensch S, Jonckers TH, Koul A, Arnoult E, Peeples ME, Roymans D, McLellan JS.
Molecular mechanism of respiratory syncytial virus fusion inhibitors.
Nat Chem Biol. 2016 Feb;12(2):87-93.
Krarup A, Truan D, Furmanova-Hollenstein P, Bogaert L, Bouchier P, Bisschop IJ, Widjojoatmodjo MN, Zahn R, Schuitemaker H, McLellan JS, Langedijk JP.
A highly stable prefusion RSV F vaccine derived from structural analysis of the fusion mechanism.
Nat Commun. 2015 Sep 3;6:8143
Gilman MS, Moin SM, Mas V, Chen M, Patel NK, Kramer K, Zhu Q, Kabeche SC, Kumar A, Palomo C, Beaumont T, Baxa U, Ulbrandt ND, Melero JA, Graham BS, McLellan JS.
Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.
PLoS Pathog. 2015 Jul 10; 11(7):e1005035.