A surface plasmon resonance assay for the binding of influenza virus hemagglutinin to its sialic acid receptor.
We have developed a sensitive microscale binding assay to study the interaction between influenza hemagglutinin and its cell surface receptor sialic acid using real-time surface plasmon resonance. The glycoprotein fetuin was bound to a carboxymethylated-Dextran sensor surface using N-hydroxysuccinimide and N-ethyl-N'-(dimethylaminopropyl) carbodiimide. Low-pH-induced BHA rosettes bind specifically to the fetuin-derivitized sensor surface, but not to an asialofetuin-derivitized sensor surface. Binding can be inhibited by preincubation of BHA rosettes with millimolar concentrations of inhibitors of the influenza hemagglutinin-sialic acid interaction. The association rate, dissociation rate, and dissociation constant for the multivalent interaction between BHA rosettes and the fetuin-derivitized sensor surface were also measured, allowing us to quantitate the tight binding achieved through the multivalent interaction between BHA rosettes and the fetuin-derivitized sensor surface.
Takemoto
D K
DK
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Skehel
J J
JJ
Wiley
D C
DC
D006389:Hemagglutinins, Viral / Q000378:metabolism*; D007700:Kinetics; D008722:Methods; D011991:Receptors, Virus / Q000378:metabolism*; D012397:Rosette Formation; D012794:Sialic Acids / Q000378:metabolism*; D013057:Spectrum Analysis; D000509:alpha-Fetoproteins / Q000737:chemistry
D006389:Hemagglutinins, Viral; D011991:Receptors, Virus; D012794:Sialic Acids; D000509:alpha-Fetoproteins