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The kinetics of the acid-induced conformational change of influenza virus haemagglutinin can be followed using 1,1'-bis(4-anilino-5-naphthalenesulphonic acid).

Abstract
1,1'-Bis(4-anilino-5-naphthalenesulphonic acid) (bis-ANS) has been shown by fluorescence spectroscopy to bind to bromelain-cleaved influenza haemagglutinin (BHA). The fluorescence intensity of 1.2 microM bis-ANS in the presence of BHA in its low-pH conformation is twenty-fold higher than in the presence of BHA in its neutral-pH conformation. The use of this probe provides a sensitive method for investigating the kinetics of the irreversible conformational change of BHA induced by low pH. At pH5.0 the reaction is described by a rapid burst followed by a double exponential increase in the fluorescence of bis-ANS, with rate constants of 5.2 +/- 0.9 x 10(-3) sec-1 and 6.7 +/- 1.9 x 10(-4) sec-1. This reaction is sensitive to the presence of tert-butylhydroquinone, an inhibitor of the conformational transition of BHA. The dependence of the reaction rate on pH indicates that the acid-induced conformational change is dependent upon the multiple protonation of the neutral-pH conformation of BHA.
AuthorsR C Bethell, N M Gray, C R Penn
JournalBiochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 206 Issue 1 Pg. 355-61 (Jan 05 1995) ISSN: 0006-291X [Print] United States
PMID7818541 (Publication Type: Journal Article)
Chemical References
  • Anilino Naphthalenesulfonates
  • Antioxidants
  • Fluorescent Dyes
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Hemagglutinins, Viral
  • Hydroquinones
  • Peptide Fragments
  • Viral Envelope Proteins
  • 5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate)
  • Bromelains
  • 2-tert-butylhydroquinone
Topics
  • Anilino Naphthalenesulfonates
  • Antioxidants (pharmacology)
  • Bromelains
  • Fluorescent Dyes
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Hemagglutinins, Viral (chemistry)
  • Hydrogen-Ion Concentration
  • Hydroquinones (pharmacology)
  • Kinetics
  • Peptide Fragments (chemistry)
  • Protein Conformation (drug effects)
  • Spectrometry, Fluorescence
  • Time Factors
  • Viral Envelope Proteins (chemistry)

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