Zn 2+-induced rearrangement of the disordered TPPP/p25 affects its microtubule assembly and GTPase activity

Ágnes Zotter, Judit Oláh, Emma Hlavanda, Andrea Bodor, András Perczel, Krisztián Szigeti, Judit Fidy, Judit Ovádi

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Abstract

Tubulin polymerization promoting protein/p25 (TPPP/p25) modulates the dynamics and stability of the microtubule system and plays crucial role in the myelination of oligodendrocytes. Here we showed by CD, fluorescence, and NMR spectroscopies that Zn 2+ is the first ligand that induces considerable rearrangement of the disordered TPPP/p25. Zinc finger motif (His 2Cys 2) (His 61-Cys 83) was identified within the flexible region of TPPP/p25 straddled by extended unstructured N- and C-terminal regions. The specific binding of the Zn 2+ to TPPP/p25 induced the formation of molten globule but not that of a well-defined tertiary structure. The Zn 2+-induced partially folded structure accommodating the zinc binding motif is localized at the single Trp 76-containing region as demonstrated by fluorescence resonance energy transfer and quenching experiments. We showed that the Zn 2+-induced change in the TPPP/p25 structure modified its interaction with tubulin and GTP coupled with functional consequences: the TPPP/p25-promoted tubulin polymerization was increased while the TPPP/p25-catalyzed GTPase activity was decreased as detected by turbidimetry and by malachite green phosphate release/ 31P NMR assays, respectively. The finding that the Zn 2+ of the bivalent cations can uniquely influence physiological relavant interactions significantly contributes to our understanding of the role of Zn 2+-related TPPP/p25 processes in the differentiation/ myelination of oligodendrocytes possessing a high-affinity Zn 2+ uptake mechanism.

Original languageEnglish
Pages (from-to)9568-9578
Number of pages11
JournalBiochemistry
Volume50
Issue number44
DOIs
Publication statusPublished - Nov 8 2011

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GTP Phosphohydrolases
Tubulin
Microtubules
Polymerization
Thermodynamic properties
Proteins
Oligodendroglia
Zinc
Nephelometry and Turbidimetry
Fluorescence Resonance Energy Transfer
Fluorescence Spectrometry
Zinc Fingers
Fluorescence spectroscopy
Guanosine Triphosphate
Nuclear magnetic resonance spectroscopy
Cations
Molten materials
Quenching
Assays
Magnetic Resonance Spectroscopy

ASJC Scopus subject areas

  • Biochemistry

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Zn 2+-induced rearrangement of the disordered TPPP/p25 affects its microtubule assembly and GTPase activity. / Zotter, Ágnes; Oláh, Judit; Hlavanda, Emma; Bodor, Andrea; Perczel, András; Szigeti, Krisztián; Fidy, Judit; Ovádi, Judit.

In: Biochemistry, Vol. 50, No. 44, 08.11.2011, p. 9568-9578.

Research output: Contribution to journalArticle

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