Reversible acetonitrile-induced inactivation/activation of thermolysin

R.V. Ulijn, A.E.M. Janssen, B.D. Moore, P.J. Halling, S.M. Kelly, N.C. Price

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Thermolysin is catalytically inactive in mixtures or 70-15% acetonitrile in aqueous buffer. Unexpectedly, dilution of the inactive enzyme with acetonitrile leads to complete recovery of the catalytic activity in a similar way to dilution with aqueous buffer. Circular dichroism and fluorescence studies of thermolysin in the same solvent mixtures reveal discontinuous changes in the overall secondary and tertiary protein structure that correlate well with the reversible differences in catalytic activity. The spectra on either side of the minimum activity point are different from each other, a fact indicating that the enzyme may be able to access two active conformations which are thermodynamically stable in different solvent environments.
LanguageEnglish
Pages1112-1116
Number of pages4
JournalChembiochem
Volume3
Issue number11
DOIs
Publication statusPublished - 4 Nov 2002

Fingerprint

Thermolysin
Dilution
Catalyst activity
Buffers
Chemical activation
Secondary Protein Structure
Enzymes
Circular Dichroism
Tertiary Protein Structure
Conformations
Fluorescence
Recovery
Proteins
acetonitrile

Keywords

  • activity studies
  • biocatalysis
  • enzymes
  • protein structures
  • solvent effects
  • Aqueous-organic media
  • alpha-chymotrypsin
  • catalytic activity
  • solvents
  • fluorescence
  • denaturation
  • mixtures
  • relevant

Cite this

Ulijn, R.V. ; Janssen, A.E.M. ; Moore, B.D. ; Halling, P.J. ; Kelly, S.M. ; Price, N.C. / Reversible acetonitrile-induced inactivation/activation of thermolysin. In: Chembiochem. 2002 ; Vol. 3, No. 11. pp. 1112-1116.
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Reversible acetonitrile-induced inactivation/activation of thermolysin. / Ulijn, R.V.; Janssen, A.E.M.; Moore, B.D.; Halling, P.J.; Kelly, S.M.; Price, N.C.

In: Chembiochem, Vol. 3, No. 11, 04.11.2002, p. 1112-1116.

Research output: Contribution to journalArticle

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T1 - Reversible acetonitrile-induced inactivation/activation of thermolysin

AU - Ulijn, R.V.

AU - Janssen, A.E.M.

AU - Moore, B.D.

AU - Halling, P.J.

AU - Kelly, S.M.

AU - Price, N.C.

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AB - Thermolysin is catalytically inactive in mixtures or 70-15% acetonitrile in aqueous buffer. Unexpectedly, dilution of the inactive enzyme with acetonitrile leads to complete recovery of the catalytic activity in a similar way to dilution with aqueous buffer. Circular dichroism and fluorescence studies of thermolysin in the same solvent mixtures reveal discontinuous changes in the overall secondary and tertiary protein structure that correlate well with the reversible differences in catalytic activity. The spectra on either side of the minimum activity point are different from each other, a fact indicating that the enzyme may be able to access two active conformations which are thermodynamically stable in different solvent environments.

KW - activity studies

KW - biocatalysis

KW - enzymes

KW - protein structures

KW - solvent effects

KW - Aqueous-organic media

KW - alpha-chymotrypsin

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KW - solvents

KW - fluorescence

KW - denaturation

KW - mixtures

KW - relevant

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