title: Antibody recognition of the pandemic H1N1 Influenza virus hemagglutinin receptor binding site.
abstract: Influenza virus is a global health concern due to its unpredictable pandemic potential. This potential threat was realized in 2009 when an H1N1 virus emerged that resembled the 1918 virus in antigenicity but fortunately was not nearly as deadly. 5J8 is a human antibody that potently neutralizes a broad spectrum of H1N1 viruses, including the 1918 and 2009 pandemic viruses. Here, we present the crystal structure of 5J8 Fab in complex with a bacterially expressed and refolded globular head domain from the hemagglutinin (HA) of the A/California/07/2009 (H1N1) pandemic virus. 5J8 recognizes a conserved epitope in and around the receptor binding site (RBS), and its HCDR3 closely mimics interactions of the sialic acid receptor. Electron microscopy (EM) reconstructions of 5J8 Fab in complex with an HA trimer from a 1986 H1 strain and with an engineered stabilized HA trimer from the 2009 H1 pandemic virus showed a similar mode of binding. As for other characterized RBS-targeted antibodies, 5J8 uses avidity to extend its breadth and affinity against divergent H1 strains. 5J8 selectively interacts with HA insertion residue 133a, which is conserved in pandemic H1 strains and has precluded binding of other RBS-targeted antibodies. Thus, the RBS of divergent HAs is targeted by 5J8 and adds to the growing arsenal of common recognition motifs for design of therapeutics and vaccines. Moreover, consistent with previous studies, the bacterially expressed H1 HA properly refolds, retaining its antigenic structure, and presents a low-cost and rapid alternative for engineering and manufacturing candidate flu vaccines.
authors.affiliation: Department of Integrative Structural and Computational Biology.
authors.forename: Minsun
authors.initials: M
authors.lastname: Hong
authors.forename: Peter S
authors.initials: PS
authors.lastname: Lee
authors.forename: Ryan M B
authors.initials: RM
authors.lastname: Hoffman
authors.forename: Xueyong
authors.initials: X
authors.lastname: Zhu
authors.forename: Jens C
authors.initials: JC
authors.lastname: Krause
authors.forename: Nick S
authors.initials: NS
authors.lastname: Laursen
authors.forename: Sung-Il
authors.initials: SI
authors.lastname: Yoon
authors.forename: Langzhou
authors.initials: L
authors.lastname: Song
authors.forename: Lynda
authors.initials: L
authors.lastname: Tussey
authors.forename: James E
authors.initials: JE
authors.lastname: Crowe
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Ward
authors.forename: Ian A
authors.initials: IA
authors.lastname: Wilson
mesh_terms: D000914:Antibodies, Viral / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity; D000956:Antigens, Viral / Q000276:immunology* / Q000378:metabolism; D001666:Binding Sites, Antibody; D018360:Crystallography, X-Ray; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007251:Influenza, Human / Q000453:epidemiology / Q000276:immunology* / Q000821:virology; D008958:Models, Molecular; D058873:Pandemics; D011487:Protein Conformation; D011991:Receptors, Virus / Q000276:immunology* / Q000378:metabolism; D014481:United States / Q000453:epidemiology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; C543345:H1N1 virus hemagglutinin; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011991:Receptors, Virus
references.citation: Immunol Rev. 2012 Nov;250(1):180-98
references.pmid: 23046130
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32
references.pmid: 20124692
references.citation: Nucleic Acids Res. 2004 Mar 19;32(5):1792-7
references.pmid: 15034147
references.citation: J Mol Biol. 1987 Sep 20;197(2):273-96
references.pmid: 3681996
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1458-63
references.pmid: 23297216
references.citation: J Virol. 2008 Jan;82(2):596-601
references.pmid: 17942553
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 2;110(1):264-9
references.pmid: 23175789
references.citation: Proc Natl Acad Sci U S A. 2012 Oct 16;109(42):17040-5
references.pmid: 23027945
references.citation: J Virol. 2011 Jan;85(2):865-72
references.pmid: 21068239
references.citation: Science. 2013 May 10;340(6133):711-6
references.pmid: 23539181
references.citation: PLoS One. 2010 Jul 22;5(7):e11694
references.pmid: 20661476
references.citation: Nature. 1981 Jan 29;289(5796):366-73
references.pmid: 7464906
references.citation: J Struct Biol. 2010 Aug;171(2):197-206
references.pmid: 20362059
references.citation: J Struct Biol. 2009 Apr;166(1):95-102
references.pmid: 19263523
references.citation: J Struct Biol. 1999 Dec 1;128(1):82-97
references.pmid: 10600563
references.citation: PLoS Curr. 2010 Mar 22;2:RRN1152
references.pmid: 20352039
references.citation: Proc Natl Acad Sci U S A. 2011 Aug 23;108(34):14216-21
references.pmid: 21825125
references.citation: J Comput Chem. 2004 Oct;25(13):1605-12
references.pmid: 15264254
references.citation: PLoS One. 2011;6(6):e20928
references.pmid: 21687743
references.citation: Vaccine. 2010 Dec 6;28(52):8268-74
references.pmid: 20969925
references.citation: Nature. 2012 Sep 27;489(7417):526-32
references.pmid: 22982990
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
references.pmid: 20057044
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references.pmid: 15299926
references.citation: Nat Struct Mol Biol. 2013 Mar;20(3):363-70
references.pmid: 23396351
references.citation: J Virol. 2012 Jan;86(2):982-90
references.pmid: 22072785
references.citation: J Struct Biol. 2009 May;166(2):205-13
references.pmid: 19374019
references.citation: J Struct Biol. 1996 Oct;116(1):237-40
references.pmid: 8812978
references.citation: J Virol. 2011 Oct;85(20):10905-8
references.pmid: 21849447
references.citation: Science. 2010 Apr 16;328(5976):357-60
references.pmid: 20339031
references.citation: Science. 2011 Aug 12;333(6044):843-50
references.pmid: 21737702
references.citation: Mol Immunol. 2008 Aug;45(14):3832-9
references.pmid: 18614234
references.citation: Protein Cell. 2010 May;1(5):459-67
references.pmid: 21203961
references.citation: J Mol Biol. 1994 May 20;238(5):777-93
references.pmid: 8182748
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: Nature. 1988 Jun 2;333(6172):426-31
references.pmid: 3374584
references.citation: Nature. 1981 Apr 23;290(5808):713-7
references.pmid: 6163993
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
references.pmid: 20124702
references.citation: PLoS Pathog. 2012;8(12):e1003067
references.pmid: 23236279
references.citation: J Struct Biol. 1996 Jan-Feb;116(1):190-9
references.pmid: 8742743
references.citation: Protein Eng. 1996 Jul;9(7):617-21
references.pmid: 8844834
references.citation: J Struct Biol. 2005 Jul;151(1):41-60
references.pmid: 15890530
references.citation: J Struct Biol. 2000 Oct;132(1):33-45
references.pmid: 11121305
references.citation: Nature. 1995 Jul 6;376(6535):92-4
references.pmid: 7596443
references.citation: J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
references.pmid: 19461840
references.citation: Vaccine. 2012 Aug 24;30(39):5761-9
references.pmid: 22796139
references.citation: Cell. 1982 Dec;31(2 Pt 1):417-27
references.pmid: 6186384
references.citation: Science. 2011 May 13;332(6031):816-21
references.pmid: 21566186
references.citation: Nat Biotechnol. 2012 May 27;30(6):543-8
references.pmid: 22634563