Helix mobility and recognition function of the rat thyroid transcription factor 1 homeodomain - Hints from 15N-NMR relaxation studies

Devrim Gümral, Luana Nadalin, Alessandra Corazza, Federico Fogolari, Giuseppe Damante, Paolo Viglino, Gennaro Esposito

    Research output: Contribution to journalArticle

    Abstract

    The backbone dynamics of the 15N-labeled homeodomain of the rat thyroid transcription factor 1 has been studied by 2D NMR spectroscopy. Longitudinal (R1) and transverse (R2) 15N relaxation rate constants and steady-state {1H}-15N NOEs were measured at 11.7 T. These data were analyzed by both the model-free formalism and the reduced spectral density mapping (RSDM) approaches. The global rotational correlation time, τm, of the thyroid transcription factor 1 homeodomain in aqueous solution at 286 K was found to be 10.51 ± 0.05 ns by model-free formalism and 9.85 ± 1.79 ns by RSDM calculation. The homogeneity of the values of the overall correlation time calculated from the individual (R2/R1) ratios suggested a good degree of isotropy of the global molecular motion, consistent with the similar global τm results obtained with the two different methods. Tyr25 was found to undergo slow conformational exchange by both methods, whereas this contribution was identified also for Lys21, Gln22, Ile38 and His52 only by RSDM. With both methods, the C-terminal fragment of helix III was found to be more flexible than the preceding N-terminal portion, with slightly different limits between rigid and mobile moieties. Additionally, Arg53 appeared to be characterized by an intermediate motional freedom between the very flexible N-terminal and C-terminal residues and the structured core of the molecule, suggesting the occurrence of a hinge point. Finally, slow-time-scale motions observed at the end of helix I, at the end of helix II and within helix III appear to be consistent with typical fraying transitions at helical C-termini.

    Original languageEnglish (US)
    Pages (from-to)435-448
    Number of pages14
    JournalFEBS Journal
    Volume275
    Issue number3
    DOIs
    StatePublished - Feb 1 2008

    Fingerprint

    Spectral density
    Nuclear magnetic resonance
    Hinges
    Nuclear magnetic resonance spectroscopy
    Rate constants
    Magnetic Resonance Spectroscopy
    Molecules
    thyroid nuclear factor 1

    Keywords

    • Backbone dynamics
    • Model-free approach
    • NMR N relaxation
    • Spectral density mapping
    • Thyroid transcription factor 1 homeodomain

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Helix mobility and recognition function of the rat thyroid transcription factor 1 homeodomain - Hints from 15N-NMR relaxation studies. / Gümral, Devrim; Nadalin, Luana; Corazza, Alessandra; Fogolari, Federico; Damante, Giuseppe; Viglino, Paolo; Esposito, Gennaro.

    In: FEBS Journal, Vol. 275, No. 3, 01.02.2008, p. 435-448.

    Research output: Contribution to journalArticle

    Gümral, Devrim ; Nadalin, Luana ; Corazza, Alessandra ; Fogolari, Federico ; Damante, Giuseppe ; Viglino, Paolo ; Esposito, Gennaro. / Helix mobility and recognition function of the rat thyroid transcription factor 1 homeodomain - Hints from 15N-NMR relaxation studies. In: FEBS Journal. 2008 ; Vol. 275, No. 3. pp. 435-448.
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    abstract = "The backbone dynamics of the 15N-labeled homeodomain of the rat thyroid transcription factor 1 has been studied by 2D NMR spectroscopy. Longitudinal (R1) and transverse (R2) 15N relaxation rate constants and steady-state {1H}-15N NOEs were measured at 11.7 T. These data were analyzed by both the model-free formalism and the reduced spectral density mapping (RSDM) approaches. The global rotational correlation time, τm, of the thyroid transcription factor 1 homeodomain in aqueous solution at 286 K was found to be 10.51 ± 0.05 ns by model-free formalism and 9.85 ± 1.79 ns by RSDM calculation. The homogeneity of the values of the overall correlation time calculated from the individual (R2/R1) ratios suggested a good degree of isotropy of the global molecular motion, consistent with the similar global τm results obtained with the two different methods. Tyr25 was found to undergo slow conformational exchange by both methods, whereas this contribution was identified also for Lys21, Gln22, Ile38 and His52 only by RSDM. With both methods, the C-terminal fragment of helix III was found to be more flexible than the preceding N-terminal portion, with slightly different limits between rigid and mobile moieties. Additionally, Arg53 appeared to be characterized by an intermediate motional freedom between the very flexible N-terminal and C-terminal residues and the structured core of the molecule, suggesting the occurrence of a hinge point. Finally, slow-time-scale motions observed at the end of helix I, at the end of helix II and within helix III appear to be consistent with typical fraying transitions at helical C-termini.",
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    AU - Gümral, Devrim

    AU - Nadalin, Luana

    AU - Corazza, Alessandra

    AU - Fogolari, Federico

    AU - Damante, Giuseppe

    AU - Viglino, Paolo

    AU - Esposito, Gennaro

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    AB - The backbone dynamics of the 15N-labeled homeodomain of the rat thyroid transcription factor 1 has been studied by 2D NMR spectroscopy. Longitudinal (R1) and transverse (R2) 15N relaxation rate constants and steady-state {1H}-15N NOEs were measured at 11.7 T. These data were analyzed by both the model-free formalism and the reduced spectral density mapping (RSDM) approaches. The global rotational correlation time, τm, of the thyroid transcription factor 1 homeodomain in aqueous solution at 286 K was found to be 10.51 ± 0.05 ns by model-free formalism and 9.85 ± 1.79 ns by RSDM calculation. The homogeneity of the values of the overall correlation time calculated from the individual (R2/R1) ratios suggested a good degree of isotropy of the global molecular motion, consistent with the similar global τm results obtained with the two different methods. Tyr25 was found to undergo slow conformational exchange by both methods, whereas this contribution was identified also for Lys21, Gln22, Ile38 and His52 only by RSDM. With both methods, the C-terminal fragment of helix III was found to be more flexible than the preceding N-terminal portion, with slightly different limits between rigid and mobile moieties. Additionally, Arg53 appeared to be characterized by an intermediate motional freedom between the very flexible N-terminal and C-terminal residues and the structured core of the molecule, suggesting the occurrence of a hinge point. Finally, slow-time-scale motions observed at the end of helix I, at the end of helix II and within helix III appear to be consistent with typical fraying transitions at helical C-termini.

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