pH-dependent binding of the fluorophore bis-ANS to influenza virus reflects the conformational change of hemagglutinin.
Binding of the fluorophore 1,1'-bis(4-anilino)naphthalene-5,5'-disulfonic acid (bis-ANS) to influenza virus A/PR 8/34 is strongly enhanced at low pH. Binding is accompanied by a significant increase in fluorescence intensity. The binding and the fluorescence increase are associated with the low-pH induced conformational change of the viral spike protein, hemagglutinin, exposing hydrophobic binding sites. The data indicate that in addition to the hydrophobic N-terminus of HA2 other hydrophobic sequences of the HA ectodomain become accessible to bis-ANS at low pH. It is shown that the time course of the fluorescence increase of bis-ANS at low pH is determined by the conformational change of HA. The application of this assay for continuously monitoring the kinetics of the structural alteration in HA is discussed and its relevance for elucidating the temporal relationship between the conformational change of HA and virus-membrane fusion is outlined.
Korte
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Institut für Biophysik, Fachbereich Biologie, Humboldt-Universität zu Berlin, Germany.
Herrmann
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D000817:Anilino Naphthalenesulfonates / Q000737:chemistry*; D004910:Erythrocyte Membrane / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006389:Hemagglutinins, Viral / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000737:chemistry / Q000648:ultrastructure*; D008561:Membrane Fusion; D011487:Protein Conformation; D014760:Viral Fusion Proteins / Q000737:chemistry
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D006389:Hemagglutinins, Viral; D014760:Viral Fusion Proteins; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate)