Effects of transketolase cofactors on its conformation and stability

Olga A. Esakova, Ludmilla E. Meshalkina, German A. Kochetov

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

In studying transketolase (TK) from Saccharomyces cerevisiae, the majority of researchers use as cofactors Mg2+ and thiamine diphosphate (ThDP) (by analogy with other ThDP-dependent enzymes), whereas the active site of native holoTK is known to contain only Ca2+. Experiments in which Mg2+ was substituted for Ca2+ demonstrated that the kinetic properties of TK varied with the bivalent cation cofactor. This led to the assumption that TK species obtained by reconstitution from apoTK and ThDP in the presence of Ca2+ or Mg2+, respectively, adopt different conformations [Selivanov, V.A., Kovina, M.V., Kochevova, N.V., Meshalkina, L.E., Kochetov, G.A., 2004. Kinetic study of the H103A mutant yeast transketolase. FEBS Letters 567, 270-274]. Analysis of far-UV circular dichroism (CD) spectra and of data, obtained using methods of thermal denaturing, differential scanning calorimetry (DSC) and tryptophan fluorescence spectroscopy, corroborated this assumption. Indeed, the ratios of secondary structure elements in the molecule of apoTK, recorded in the presence of Ca 2+ or Mg2+, respectively, turned out to be different. The two forms of the holoenzyme, obtained by reconstitution from apoTK and ThDP in the presence of Ca2+ or Mg2+, respectively, also differed in stability: the holoenzyme was more stable in the presence of Ca2+ than Mg2+.

Original languageEnglish (US)
Pages (from-to)8-13
Number of pages6
JournalLife Sciences
Volume78
Issue number1
DOIs
StatePublished - Nov 19 2005

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Transketolase
Thiamine Pyrophosphate
Conformations
Holoenzymes
Yeast
Kinetics
Fluorescence Spectrometry
Differential Scanning Calorimetry
Fluorescence spectroscopy
Circular Dichroism
Tryptophan
Saccharomyces cerevisiae
Cations
Differential scanning calorimetry
Catalytic Domain
Hot Temperature
Yeasts
Research Personnel
Molecules
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Esakova, Olga A. ; Meshalkina, Ludmilla E. ; Kochetov, German A. / Effects of transketolase cofactors on its conformation and stability. In: Life Sciences. 2005 ; Vol. 78, No. 1. pp. 8-13.
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Effects of transketolase cofactors on its conformation and stability. / Esakova, Olga A.; Meshalkina, Ludmilla E.; Kochetov, German A.

In: Life Sciences, Vol. 78, No. 1, 19.11.2005, p. 8-13.

Research output: Contribution to journalArticle

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