Cold inactivation and dissociation into dimers of Escherichia coli tryptophanase and its W330F mutant form

Tali Erez, Garik Ya Gdalevsky, Yuri M. Torchinsky, Robert S. Phillips, Abraham H. Parola

Research output: Contribution to journalArticlepeer-review

20 Scopus citations


The kinetics and mechanism of reversible cold inactivation of the tetrameric enzyme tryptophanase have been studied. Cold inactivation is shown to occur slowly in the presence of K+ ions and much faster in their absence. The W330F mutant tryptophanase undergoes rapid cold inactivation even in the presence of K+ ions. In all cases the inactivation is accompanied by a decrease of the coenzyme 420-nm CD and absorption peaks and a shift of the latter peak to shorter wavelengths. The spectral changes and the NaBH4 test indicate that cooling of tryptophanase leads to breaking of the internal aldimine bond and release of the coenzyme. HPLC analysis showed that the ensuing apoenzyme dissociates into dimers. The dissociation depends on the nature and concentration of anions in the buffer solution. It readily occurs at low protein concentrations in the presence of salting-in anions Cl-, NO3- and I-, whereas salting-out anions, especially HPO42-, hinder the dissociation. K+ ions do not influence the dissociation of the apoenzyme, but partially protect holotryptophanase from cold inactivation. Thus, the two processes, cold inactivation of tryptophanase and dissociation of its apoform into dimers exhibit different dependencies on K+ ions and anions. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)365-372
Number of pages8
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Issue number2
StatePublished - 19 May 1998


  • Cold lability
  • Mutant enzyme
  • Protein dissociation
  • Pyridoxal phosphate
  • Tryptophanase

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology


Dive into the research topics of 'Cold inactivation and dissociation into dimers of Escherichia coli tryptophanase and its W330F mutant form'. Together they form a unique fingerprint.

Cite this