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2642613264
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2O. Incubation of this protein with (E)-1 and R2 generated an EPR spectrum identical to that shown in Figure 1C (data not shown). This result indicates that both hyperfine couplings in the radical are substrate-derived, consistent with Scheme 3.
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2642710508
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note
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• species. Its spectral position is sensitive to the protonation (or hydrogen-bonding) state of the phenolic oxygen, while the width of the peak is determined by hyperfine splittings to ring ortho and β protons and, therefore, sensitive to rotational motion of the aromatic ring. Inspection of Figure 5A,B reveals that the predominant tyrosyl radical species contributing to the t = 30 s spectrum is a tyrosyl radical structurally very similar to the tyrosyl radical at t = 0 s, since this downfield peak has essentially the same spectral position and width. Thus, the spectral subtraction methodology employed here is valid within experimental uncertainty.
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26
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2642711331
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2642704657
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note
-
In Scheme 5, the conjugate addition is arbitrarily drawn to occur from the α face (bottom face) of the enone. Addition from the β face would generate the epimers at C6′ of the structures in Scheme 5, but this would still lead to a 1:1 mixture of protonated and deuterated allyl radical species.
-
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-
-
35
-
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2642707955
-
-
note
-
15 Thus, by process of elimination, formation of 18 (Scheme 6) would require that the thiyl radical at Cys439 adds to enone 4 from its β face.
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0011235203
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