Glycoconjugate Journal

, Volume 30, Issue 7, pp 709–716 | Cite as

Binding kinetics of sulfatide with influenza A virus hemagglutinin

  • Tadanobu Takahashi
  • Sawako Kawagishi
  • Midori Masuda
  • Takashi SuzukiEmail author


Association of a sulfated galactosyl ceramide, sulfatide, with the viral envelope glycoprotein hemagglutinin (HA) delivered to the cell surface is required for influenza A virus (IAV) replication through efficient translocation of the newly synthesized viral nucleoprotein from the nucleus to the cytoplasm. To determine whether the ectodomain of HA can bind to sulfatide, a secreted-type HA (sHA), in which the transmembrane region and cytoplasmic tail were deleted, was generated by using a baculovirus expression system. The receptor binding ability and antigenic structure of sHA were evaluated by a hemagglutination assay, solid-phase binding assay and hemagglutination inhibition assay. sHA showed subtype-specific antigenicity and binding ability to both sulfatide and gangliosides. Kinetics of sHA binding to sulfatide and GD1a was demonstrated by quartz crystal microbalance (QCM) analysis. QCM analysis showed that the sHA bound with the association rate constant (k on) of 1.41 × 104 M−1 sec−1, dissociation rate constant (k off) of 2.03 × 10−4 sec−1 and K d of 1.44 × 10−8 M to sulfatide immobilized on a sensor chip. The k off values of sHA were similar for sulfatide and GD1a, whereas the k on value of sHA binding to sulfatide was 2.56-times lower than that of sHA binding to GD1a. The results indicate that sulfatide directly binds to the ectodomain of HA with high affinity.


Baculovirus Binding affinity Influenza virus Hemagglutinin Sulfatide Quartz crystal microbalance analysis 



This work was supported by the Global COE Program from the Japan Society for the Promotion of Science and by a grant-in-aid for Scientific Research of Young Scientists B (20790357), MEXT/JSPS KAKENHI Grant Number (C; 23590549), Grant from Mizutani Foundation for Glycoscience, Grant-in-Aid from the Tokyo Biochemical Research Foundation, Grant-in-Aid from the Hamamatsu Scientific Research Foundation, Grant-in-aid from Takeda Science Foundation, Sasakawa Scientific Research Grant from the Japan Science Society, Grant-in-Aid from the Research Foundation for Pharmaceutical Sciences, Grant-in-aid from Hokuto Foundation for Bioscience, research grant of the Institute for Fermentation, Osaka and Adaptable and Seamless Technology Transfer Program (A-step) through Target-driven R&D, JST


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Tadanobu Takahashi
    • 1
  • Sawako Kawagishi
    • 1
  • Midori Masuda
    • 1
  • Takashi Suzuki
    • 1
    Email author
  1. 1.Department of Biochemistry, School of Pharmaceutical Sciences and Global COE Program for Innovation in Human Health SciencesUniversity of ShizuokaShizuokaJapan

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