Physico-chemical characterization of a recombinant cytoplasmic form of lysine: N6-hydroxylase.
A recombinant cytoplasmic preparation of lysine: N6-hydroxylase, IucD398, with a deletion of 47 amino acids at the N-terminus, was purified to homogeneity. IucD398 is capable of N-hydroxylation of L-lysine upon supplementation with FAD and NADPH. The enzyme is stringently specific with L-lysine and (S)-2-aminoethyl-L-cysteine serving as substrates. Protonophores, FCCP and CCCP, as well as cinnamylidene, have been found to serve as potent inhibitors of lysine: N6-hydroxylation by virtue of their ability to interfere in the reduction of the flavin cofactor.
Thariath
A M
AM
Guelph Waterloo Centre for Graduate Work in Chemistry, Department of Chemistry, University of Waterloo, Ontario, Canada.
Fatum
K L
KL
Valvano
M A
MA
Viswanatha
T
T
D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D003593:Cytoplasm / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology; D005182:Flavin-Adenine Dinucleotide; D006861:Hydrogen Peroxide / Q000737:chemistry; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D009249:NADP; D011994:Recombinant Proteins / Q000737:chemistry; D013379:Substrate Specificity
D011994:Recombinant Proteins; D005182:Flavin-Adenine Dinucleotide; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D009249:NADP; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D006861:Hydrogen Peroxide; D006899:Mixed Function Oxygenases; C047196:lysine N-epsilon hydroxylase