Alkaline unfolding and salt-induced folding of yeast alcohol dehydrogenase under high pH conditions

L. E. Wei-Ping, S. X. Yan, L. I. Sen, Haining Zhong, Hai Meng Zhou

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The conformational changes of yeast alcohol dehydrogenase during unfolding at alkaline pH have been followed by fluorescence emission and circular dichroism spectra. A result of comparison of inactivation and conformational changes shows that much lower values of alkaline pH are required to bring about inactivation than significant conformational change of the enzyme molecule. At pH 9.5, although the enzyme has been completely inactivated, no marked conformational changes can be observed. Even at pH 12, the apparently fully unfolded enzyme retains some ordered secondary structure. After removal of Zn2+ from the enzyme molecule, the conformational stability decreased. At ph 12 by adding the salt, the relatively unfolded state of denatured enzyme changes into a compact conformational state by hydrophobic collapsing. Folded states induced by salt bound ANS strongly, indicating the existence of increased hydrophobic surface. More extensive studies showed that although apo-YADH and holo-YADH exhibited similar behavior, the folding cooperative ability of apo-enzyme was lower than that of holo-enzyme. The above results suggest that the zinc ion plays an important role in helping the folding of YADH and in stabilizing its native conformation.

Original languageEnglish (US)
Pages (from-to)484-490
Number of pages7
JournalInternational Journal of Peptide and Protein Research
Volume47
Issue number6
StatePublished - 1996
Externally publishedYes

Fingerprint

Alcohol Dehydrogenase
Salts
Enzymes
Molecules
Aptitude
Circular Dichroism
Cooperative Behavior
Conformations
Zinc
Fluorescence
Ions

Keywords

  • alcohol dehydrogenase
  • alkaline unfolding
  • conformation
  • folding

ASJC Scopus subject areas

  • Biochemistry

Cite this

Alkaline unfolding and salt-induced folding of yeast alcohol dehydrogenase under high pH conditions. / Wei-Ping, L. E.; Yan, S. X.; Sen, L. I.; Zhong, Haining; Zhou, Hai Meng.

In: International Journal of Peptide and Protein Research, Vol. 47, No. 6, 1996, p. 484-490.

Research output: Contribution to journalArticle

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AU - Sen, L. I.

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AU - Zhou, Hai Meng

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N2 - The conformational changes of yeast alcohol dehydrogenase during unfolding at alkaline pH have been followed by fluorescence emission and circular dichroism spectra. A result of comparison of inactivation and conformational changes shows that much lower values of alkaline pH are required to bring about inactivation than significant conformational change of the enzyme molecule. At pH 9.5, although the enzyme has been completely inactivated, no marked conformational changes can be observed. Even at pH 12, the apparently fully unfolded enzyme retains some ordered secondary structure. After removal of Zn2+ from the enzyme molecule, the conformational stability decreased. At ph 12 by adding the salt, the relatively unfolded state of denatured enzyme changes into a compact conformational state by hydrophobic collapsing. Folded states induced by salt bound ANS strongly, indicating the existence of increased hydrophobic surface. More extensive studies showed that although apo-YADH and holo-YADH exhibited similar behavior, the folding cooperative ability of apo-enzyme was lower than that of holo-enzyme. The above results suggest that the zinc ion plays an important role in helping the folding of YADH and in stabilizing its native conformation.

AB - The conformational changes of yeast alcohol dehydrogenase during unfolding at alkaline pH have been followed by fluorescence emission and circular dichroism spectra. A result of comparison of inactivation and conformational changes shows that much lower values of alkaline pH are required to bring about inactivation than significant conformational change of the enzyme molecule. At pH 9.5, although the enzyme has been completely inactivated, no marked conformational changes can be observed. Even at pH 12, the apparently fully unfolded enzyme retains some ordered secondary structure. After removal of Zn2+ from the enzyme molecule, the conformational stability decreased. At ph 12 by adding the salt, the relatively unfolded state of denatured enzyme changes into a compact conformational state by hydrophobic collapsing. Folded states induced by salt bound ANS strongly, indicating the existence of increased hydrophobic surface. More extensive studies showed that although apo-YADH and holo-YADH exhibited similar behavior, the folding cooperative ability of apo-enzyme was lower than that of holo-enzyme. The above results suggest that the zinc ion plays an important role in helping the folding of YADH and in stabilizing its native conformation.

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