Essential role of the metal-ion in the IPM-assisted domain closure of 3-isopropylmalate dehydrogenase

Éva Gráczer, Peter V. Konarev, Tamás Szimler, András Bacsó, Adrienn Bodonyi, Dmitri I. Svergun, P. Závodszky, M. Vas

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

X-ray structures of 3-isopropylmalate dehydrogenase (IPMDH) do not provide sufficient information on the role of the metal-ion in the metal-IPM assisted domain closure. Here solution studies were carried out to test its importance. Small-angle X-ray scattering (SAXS) experiments with the Thermus thermophilus enzyme (complexes with single substrates) have revealed only a very marginal (0-5%) extent of domain closure in the absence of the metal-ion. Only the metal-IPM complex, but neither the metal-ion nor the free IPM itself, is efficient in stabilizing the native protein conformation as confirmed by denaturation experiments with Escherichia coli IPMDH and by studies of the characteristic fluorescence resonance energy transfer (FRET) signal (from Trp to bound NADH) with both IPMDHs. A possible atomic level explanation of the metal-effect is given.

Original languageEnglish
Pages (from-to)3297-3302
Number of pages6
JournalFEBS Letters
Volume585
Issue number20
DOIs
Publication statusPublished - Oct 20 2011

Fingerprint

3-Isopropylmalate Dehydrogenase
Metal ions
Metals
Ions
Denaturation
Metal complexes
X ray scattering
NAD
Escherichia coli
X-Rays
Conformations
Thermus thermophilus
Experiments
Fluorescence Resonance Energy Transfer
Protein Conformation
Coordination Complexes
X rays
Substrates
Enzymes
Proteins

Keywords

  • 3-Isopropylmalate dehydrogenase
  • Domain movement
  • Small-angle
  • Substrate and metal-effect
  • X-ray scattering

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology

Cite this

Gráczer, É., Konarev, P. V., Szimler, T., Bacsó, A., Bodonyi, A., Svergun, D. I., ... Vas, M. (2011). Essential role of the metal-ion in the IPM-assisted domain closure of 3-isopropylmalate dehydrogenase. FEBS Letters, 585(20), 3297-3302. https://doi.org/10.1016/j.febslet.2011.09.013

Essential role of the metal-ion in the IPM-assisted domain closure of 3-isopropylmalate dehydrogenase. / Gráczer, Éva; Konarev, Peter V.; Szimler, Tamás; Bacsó, András; Bodonyi, Adrienn; Svergun, Dmitri I.; Závodszky, P.; Vas, M.

In: FEBS Letters, Vol. 585, No. 20, 20.10.2011, p. 3297-3302.

Research output: Contribution to journalArticle

Gráczer É, Konarev PV, Szimler T, Bacsó A, Bodonyi A, Svergun DI et al. Essential role of the metal-ion in the IPM-assisted domain closure of 3-isopropylmalate dehydrogenase. FEBS Letters. 2011 Oct 20;585(20):3297-3302. https://doi.org/10.1016/j.febslet.2011.09.013
Gráczer, Éva ; Konarev, Peter V. ; Szimler, Tamás ; Bacsó, András ; Bodonyi, Adrienn ; Svergun, Dmitri I. ; Závodszky, P. ; Vas, M. / Essential role of the metal-ion in the IPM-assisted domain closure of 3-isopropylmalate dehydrogenase. In: FEBS Letters. 2011 ; Vol. 585, No. 20. pp. 3297-3302.
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