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

doi:10.1016/S0167-4838(00)00212-0

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 journal › Article › peer-review

Abstract

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 language	English
Pages (from-to)	133-142
Number of pages	10
Journal	Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology
Volume	1544
Issue number	1-2
DOIs	https://doi.org/10.1016/S0167-4838(00)00212-0
Publication status	Published - 12 Jan 2001

Keywords

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

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10.1016/S0167-4838(00)00212-0

Cite this

Hanefeld, U., Stranzl, G., Straathof, A. J. J., Heijnen, J. J., Bergmann, A., Mittelbach, R., Glatter, O., & Kratky, C. (2001). Electrospray ionization mass spectrometry, circular dichroism and SAXS studies of the (S)-hydroxynitrile lyase from Hevea brasiliensis. Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology, 1544(1-2), 133-142. https://doi.org/10.1016/S0167-4838(00)00212-0

Electrospray ionization mass spectrometry, circular dichroism and SAXS studies of the (S)-hydroxynitrile lyase from Hevea brasiliensis. / Hanefeld, Ulf; Stranzl, Gudrun; Straathof, Adrie J.J. et al.
In: Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology, Vol. 1544, No. 1-2, 12.01.2001, p. 133-142.

Research output: Contribution to journal › Article › peer-review

Hanefeld, U, Stranzl, G, Straathof, AJJ, Heijnen, JJ, Bergmann, A, Mittelbach, R, Glatter, O & Kratky, C 2001, 'Electrospray ionization mass spectrometry, circular dichroism and SAXS studies of the (S)-hydroxynitrile lyase from Hevea brasiliensis', Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology, vol. 1544, no. 1-2, pp. 133-142. https://doi.org/10.1016/S0167-4838(00)00212-0

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abstract = "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.",

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

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AU - Hanefeld, Ulf

AU - Stranzl, Gudrun

AU - Straathof, Adrie J.J.

AU - Heijnen, Joseph J.

AU - Bergmann, Alexander

AU - Mittelbach, Rainer

AU - Glatter, Otto

AU - Kratky, Christoph

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N2 - 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.

AB - 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.

KW - Circular dichroism

KW - Cyanogenesis

KW - Electrospray ionization mass spectrometry

KW - Enzyme activity

KW - Hydroxynitrile lyase

KW - Oxynitrilase

KW - Protein conformation

KW - Small angle X-ray scattering

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JO - Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology

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IS - 1-2

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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