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Synthesis of 13 C, 15 N‐enriched α‐dicarbonyl model adducts to determine the utility of 13 c and 15 N NMR for studying mechanism‐based inactivation of cytochromes P‐450 by substituted dichloroacetamides
Author(s) -
Weiner Scott J.,
Holl Susan M.,
Covey Douglas F.
Publication year - 1994
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/mrc.1260320212
Subject(s) - chemistry , adduct , nucleophile , cysteine , serine , lysine , threonine , stereochemistry , tyrosine , organic chemistry , enzyme , amino acid , biochemistry , catalysis
Abstract To determine the feasibility of using NMR to study the inactivation of cytochromes P‐450 by dichloroacetamide‐containing mechanism‐based inactivators, 13 C, 15 N‐enriched compounds were synthesized, modeling adducts between the nucleophilic side‐chains of cysteine, lysine, serine (threonine) and tyrosine and the α‐aminooxoacyl chloride that presumably results from enzymatic conversion of a dichloroacetamide‐containing inhibitor. 13 C chemical shifts of the carbonyls attached to the nucleophiles easily distinguish the model cysteine adduct (δ 192.3) from the other adducts (δ 159.6–161.8). Although the other adducts cannot be distinguished at this carbon, the carbonyl attached to the 15 N two bonds away can distinguish the lysine mimic (δ 161.2 or 161.3) from the serine (threonine) and tyrosine mimics (δ 156.1–158.1). These assignments were made by comparing 15 N/ 13 C J values and were confirmed by solid‐state rotational echo double resonance (REDOR) studies. Finally, aminooxo‐ 15 N chemical shifts three bonds removed from the nucleophile can distinguish adducts formed to lysine (δ 108.3), cysteine (δ 104.8) and oxygen‐containing side‐chains (δ 112.8–113.9).