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Abbreviations used: HO, heme oxygenase, hHO, human heme oxygenase; rHO, rat heme oxygenase; HmuO, C. diphtheriae heme oxygenase; DMDH, 2,4-dimethyldeutrohemein; PH, protohemin: HSQC, heteronuclear single quantum coherence; NOE, nuclear Overhauser effect; NOESY, 2D nuclear Overhauser spectroscopy; TOCSY, 2D total correlation spectroscopy; DSS, 2,2-dimethyl-2-silapentane-5-sulfonate; FID, free-induction decay; TROSY, transverse relaxation-optimized spectroscopy.
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TOCSY spectra exhibited significantly more cross-peaks for hHO-DMDH-CN than hHO-PH-CN.25 However, the discovery that the symmetric heme, DMDH, yields not only the expected single species with respect to heme orientation disorder23.24 but also exhibited narrower lines due to faster interconversion of the dynamic structural heterogeneity was made at the same times as the [15N]-hNO and its PH complex was being prepared. At this time, an effective and quantitative route, without unacceptable losses, to replacing PH with DMDH in the pocket of the [2015N]hHO-PH-CN sample has not been designed but is under study.
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