Backbone Dynamics of the Bacillus subtilis Glucose Permease IIA Domain Determined from 15N NMR Relaxation Measurements

Martin J. Stone, Peter E. Wright, Wayne J. Fairbrother, Arthur G. Palmer, Jonathan Reizer, Milton H. Saier

Research output: Contribution to journalArticleResearchpeer-review

223 Citations (Scopus)


The backbone dynamics of the uniformly 15N-labeled IIA domain of the glucose permease of Bacillus subtilis have been characterized using inverse-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation time constants and steady-state {1H}-15N NOEs were measured, at a spectrometer proton frequency of 500 MHz, for 137 (91%) of the 151 protonated backbone nitrogens. These data were analyzed by using a model-free dynamics formalism to determine the generalized order parameter (S2), the effective correlation time for internal motions (Te), and 15N exchange broadening contributions (Rex) for each residue, as well as the overall molecular rotational correlation time (Tm). The T1 and T2 values for most residues were in the ranges 0.45-0.55 and 0.11-0.15 s, respectively; however, a small number of residues exhibited significantly slower relaxation. Similarly, {1H}-15N NOE values for most residues were in the range 0.72-0.80, but a few residues had much smaller positive NOEs and some exhibited negative NOEs. The molecular rotational correlation time was 6.24 ± 0.01 ns; most residues had order parameters in the range 0.75-0.90 and Te values of less than ca. 25 ps. Residues found to be more mobile than the average were concentrated in three areas: the N-terminal residues (1-13), which were observed to be highly disordered; the loop from P25 to D41, the apex ofwhich is situated adjacent to the active site and may have a role in binding to other proteins; and the region from A146 to S149. All mobile residues occurred in regions close to termini, in loops, or in irregular secondary structure.

Original languageEnglish
Pages (from-to)4394-4406
Number of pages13
Issue number18
Publication statusPublished - 1992
Externally publishedYes

Cite this