Electrospray ionization mass spectrometry, circular dichroism and SAXS studies of the (S)-hydroxynitrile lyase from Hevea brasiliensis

Ulf Hanefeld*, Gudrun Stranzl, Adrie J.J. Straathof, Joseph J. Heijnen, Alexander Bergmann, Rainer Mittelbach, Otto Glatter, Christoph Kratky

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


We report on experiments pertaining to solution properties of the (S)-hydroxynitrile lyase from Hevea brasiliensis (HbHNL). Small angle X-ray scattering unequivocally established the enzyme to occur in solution as a dimer, presumably of the same structure as in the crystal. The acid induced, irreversible deactivation of HbHNL was examined by electrospray ionization mass spectrometry (ESI-MS), circular dichroism (CD) and by measuring the enzyme activity. The deactivation is paralleled by an unfolding of the enzyme. ESI-MS of this 30 000 Da per monomer heavy protein demonstrated that unfolding took place in several stages which are paralleled by a decrease in enzyme activity. Unfolding can also be observed by CD spectroscopy, and there is a clear correlation between enzyme activity and unfolding as detected by ESI-MS and CD.

Original languageEnglish
Pages (from-to)133-142
Number of pages10
JournalBiochimica et Biophysica Acta / Protein Structure and Molecular Enzymology
Issue number1-2
Publication statusPublished - 12 Jan 2001


  • Circular dichroism
  • Cyanogenesis
  • Electrospray ionization mass spectrometry
  • Enzyme activity
  • Hydroxynitrile lyase
  • Oxynitrilase
  • Protein conformation
  • Small angle X-ray scattering

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

Cite this