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. 2010 Jul 7;99(1):273–283. doi: 10.1016/j.bpj.2010.04.002

Figure 2.

Figure 2

15N NMR line broadening evidences of millisecond fluctuations of MMP-12(E219A) (left panels) and MMP-3 catalytic domain (right panels) at 26°C and 61 MHz. (a and b) Relaxation-compensated TROSY-CPMG measurements of ΔR2 = R2(11 ms)–R2(1.7 ms) are plotted, where 11 and 1.67 ms are the spacings between π-pulses of the CPMG trains. (c and d) Spin-spin relaxation rate constant R2 was measured using an HSQC-based sequence with 1.2 ms between π-pulses (26). (Dashed lines) Thresholds above which the excess line broadening is considered significant. The thresholds are drawn 2 s−1 above the mean R2 after fine filtering-out of residues with internal motions (Table S2). Secondary structure is marked as in Fig. 1. (e and f) The structures of MMP-12(E219A) and MMP-3 catalytic domain, respectively, are depicted with red/lighter shading of bonds of residues with exchange broadening at 61 MHz, reproducing that at 81 MHz (Fig. 1). Blue/darker shading indicates bonds supplementary to 81-MHz data that did not quantify the broadening.