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AN 125 I‐LABELED N 6 ‐SUBSTITUTED AZIDO ANALOG OF NAD + FOR THE PHOTOAFFINITY LABELING OF NAD + ‐LINKED ENZYMES
Author(s) -
Chen PeiLin,
Ensor Charles Mark,
Tai HsinHsiung
Publication year - 1994
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1994.tb05133.x
Subject(s) - nad+ kinase , chemistry , nicotinamide adenine dinucleotide , photoaffinity labeling , enzyme , stereochemistry , cofactor , dissociation constant , dehydrogenase , nicotinamide mononucleotide , glycerol 3 phosphate dehydrogenase , glutamate dehydrogenase , biochemistry , nicotinamide , binding site , glutamate receptor , receptor
125 I‐ N 6 ‐(N‐[6‐N‐{5‐iodo‐4‐azidosalicyl}‐aminohexyl]‐aminocarbamoylmethyl)‐nicotinamide adenine dinucleotide ( 125 I‐ N 6 ‐I‐ASA‐AH‐NAD + ) was synthesized by coupling N 6 ‐([6‐aminohexyl]‐carbamoylmethyl)‐NAD + with 4‐azidosalicylic acid N‐hydroxysuccinimide ester followed by radioiodination. The utility of 125 I‐N 6 ‐I‐ASA‐AH‐NAD + as an effective site‐directed photoprobe was demonstrated by the photolabeling of both glutamate dehydro‐genase and 15‐hydroxyprostaglandin dehydrogenase. Both enzymes can be saturated with labeled probe with apparent dissociation constants comparable to those reported for NAD + . Photoincorporation of the probe into both enzymes was found to be protected specifically by NAD + . These results indicate that 125 I‐ N 6 ‐I‐ASA‐AH‐NAD + can be a specific photoprobe for NAD + ‐linked enzymes.